Mapping quantitative trait loci (QTLs) underlying seed vitamin E content in soybean with main,epistatic and QTL × environment effects

Soybean (Glycine max (L.) Merr.) seed contains small amounts of tocopherol, a non‐enzymatic antioxidant known as lipid‐soluble vitamin E (VE). Dietary VE contributes to a decreased risk of chronic diseases in humans and has several beneficial effects on resistance to stress in plants, and increasing VE content is an important breeding goal for increasing the nutritional value of soybean. In this study, quantitative trait loci (QTLs) underlying VE content with main, epistatic and QTL × environment effects were identified in a population of F_5 : 6 recombinant inbred lines from a cross between ‘Hefeng 25’ (a low‐VE cultivar) and ‘OAC Bayfield’ (a high‐VE cultivar). A total of 18 QTLs were detected that showed additive main effects (a) and/or additive × environment interaction effects (ae) in different environments. Moreover, 19 epistatic pairs of QTLs were found to be associated with α‐tocopherol (α‐Toc), γ‐tocopherol (γ‐Toc), δ‐tocopherol (δ‐Toc) and total VE (TE) contents. The QTLs identified in multienvironments could provide more information about QTL by environment interactions and could be useful for the marker‐assistant selection of soybean cultivars with high seed VE contents.     

Molecular mapping of soybean seed tocopherols in the cross ‘OAC Bayfield’ × ‘OAC Shire’

Soybean (Glycine max [L.] Merr.) is cultivated primarily for its protein and oil in the seed. In addition, soybean seeds contain nutraceutical compounds such as tocopherols (vitamin E), which are powerful antioxidants with health benefits. The objective of this study was to identify molecular markers linked to quantitative trait loci (QTL) that affect accumulation of soybean seed tocopherols. A recombinant inbred line (RIL) population derived from the cross ‘OAC Bayfield’ × ‘OAC Shire’ was grown in three locations over 2 years. A total of 151 SSR markers were polymorphic of which a one‐way analysis of variance identified 42 markers whereas composite interval mapping identified 26 markers linked to tocopherol QTL across 17 chromosomes. Individual QTL explained from 7% to 42% of the total phenotypic variation. Significant two‐locus epistatic interactions were identified for a total of 122 combinations in 2009 and 152 in 2010. The multiple‐locus models explained 18.4–72.2% of the total phenotypic variation. The reported QTL may be used in marker‐assisted selection (MAS) to develop high tocopherol soybean cultivars.     

Novel variation for the tocopherol profile in a sunflower created by mutagenesis and recombination

α‐Tocopherol is the main tocopherol in sunflower seeds (>90%). Because it exerts a weak antioxidant action in vitro, its partial replacement by other tocopherols is an important breeding objective in this crop. The objective of this research was to develop novel tocopherol profiles in sunflower through mutagenesis and genetic recombination. Seeds of four ‘Peredovik’ accessions were used for chemical mutagenesis with ethyl methane sulfonate (EMS). Single‐seed screening in the M₂ generation resulted in two M₂ seeds, derived from different M₁ plants, with increased γ‐tocopherol contents of 19.2% and 96.7%, respectively. M₃ progeny from the M₂ seed with the 96.7% content bred true for high c‐tocopherol content, containing more than 90%γ‐tocopherol. M₃ progeny from the M₂ seed with only 19.2%γ‐tocopherol segregated in a range from 0 to 84.6%. Selection for high c‐tocopherol content produced an M_4:5 line, designated IAST‐1, with a stable high concentration of γ‐tocopherol. Crosses between IAST‐1 and T589, with an increased b‐tocopherol content, produced F₂ segregants with trans‐gressive levels of up to 77%β‐tocopherol or up to 68% d‐tocopherol. Both novel tocopherol profiles were confirmed in the F₃ generation.     

Characterization of a γ-tocopherol methyltransferase mutant gene in wild (Carthamus oxyacanthus M. Bieb.) and cultivated safflower (C. tinctorius L.)

Safflower (Carthamus tinctorius L.) seeds contain a high proportion of tocopherols (>90 %) in the α-tocopherol form. A mutant with a high concentration of γ-tocopherol (>85 %) was identified in germplasm of wild safflower (Carthamus oxyacanthus M. Bieb.) that showed strong introgression of C. tinctorius, which allowed selection of individuals of both species with high concentrations of either α- or γ-tocopherol. The trait is controlled by a γ-tocopherol methyltransferase (γ-TMT) locus. The objective of this research was to identify γ-TMT sequence mutations associated with the high γ-tocopherol trait. Full length genomic and cDNA sequences of the γ-TMT gene were obtained from plants of C. tinctorius and C. oxyacanthus with both tocopherol profiles. Sequences from high γ-tocopherol plants showed an 11 bp deletion in exon 6 of the γ-TMT gene that disrupted the reading frame and created a premature stop codon, resulting in a predicted protein with a drastically altered amino acid sequence downstream the frameshift site. The data suggested that the frameshift mutation was underlying the γ-TMT loss of function mutant allele that determines the high γ-tocopherol phenotype. The characterized sequence change of 11 bp deletion could be used directly as a functional marker for introgression of the high γ-tocopherol trait into elite safflower cultivars.     

Identification of quantitative trait loci and candidate genes controlling the tocopherol synthesis pathway in soybean (Glycine max)

A recombinant inbred line (RIL) population was used to identify quantitative trait loci (QTLs) and their candidate genes controlling the tocopherol (Toc) synthesis pathway. The RIL population was cultivated in field conditions in 3 years. A genetic map constructed using 1624 DNA markers was used for QTL analysis. We identified 22 QTLs for seed tocopherol contents and their ratios, of which two QTL clusters on chromosomes (Chr) 9 and 14 exerted consistent large effects on tocopherol composition across the 3 years. The QTL cluster localized on Chr 9 might correspond to γ-TMT3, which controls the conversion of γ-Toc into α-Toc. The QTL cluster localized on Chr 14 was novel, which might regulate the conversion of MPBQ (a precursor of δ-Toc) into DMPBQ (the precursor of γ-Toc). The effect of the QTL cluster on Chr 14 was validated in a pair of near isogenic lines, and its candidate gene was mined. The identified QTLs and their candidate genes might be used in breeding programmes to improve α-Toc content in soybean seeds.     

Effects of high oleic acid soybean on seed yield,protein and oil contents,and seed germination revealed by near‐isogeneic lines

Typical soybean oil is composed of palmitic, stearic, oleic, linoleic and linolenic acids. High oleic acid content in soybean seed is a key compositional trait that improves oxidative stability and increases oil functionality and shelf life. Using a marker‐assisted selection method, near‐isogenic lines (NILs) of G00‐3213 for the high oleic trait were developed and yield tested. These NILs have various combinations of FAD2‐1A and FAD2‐1B alleles that were derived from the same backcrossing populations. The results indicated that G00‐3213 NILs with both homozygous mutant FAD2‐1A and FAD2‐1B alleles produced an average of 788 g/kg oleic acid content. The results also demonstrated that possessing these mutant alleles did not cause a yield reduction. Furthermore, seed germination tests across 12 temperatures (12.8–32.0°C) showed that modified seed composition for oleic acid in general did not have a major impact on seed germination. However, there was a possible reduction in seed germination vigour when high oleic seeds are planted in cold soil. The mutant FAD2‐1A and FAD2‐1B alleles did not hinder either seed or plant development.     

Identification of the QTL underlying the vitamin E content of soybean seeds

Vitamin E (VE) is an important antioxidant supplement for human health. Soybean seed extracts are the main source of VE supplements. Therefore, increasing the VE content of soybean seeds is important issue in breeding programmes. To detect quantitative trait loci (QTL) associated with VE in soybean seeds, 238 F_6:7 recombinant inbred lines (RILs) were created by crossing a high VE cultivar, ‘Beifeng 9’, with a low VE cultivar, ‘Freeborn’. A genetic map was constructed using 218 polymorphic simple sequence repeat (SSR) markers. Composite interval mapping analysis detected 66 QTLs for contents of individual and total VE, 21 for α‐tocopherol, seventeen for γ‐tocopherol, thirteen for δ‐tocopherol and fifteen for total VE. The QTLs were located on chromosomes 9, 10, 15, 18 and 19. Phenotypic variance underlain by each QTL ranged from 2.4% to 32.6%. Two major QTLs (BARCSOYSSR_10_1140–BARCSOYSSR_10_1188 and BARCSOYSSR_15_0855 to BARCSOYSSR_15_0887) associated with α‐Toc, γ‐Toc, δ‐Toc and total VE contents were mapped on chromosomes 10 and 15. They explained 12.0% and 32.6% of phenotypic variance for α‐Toc; 5.5% and 13.0% for γ‐Toc; 6.6% and 23.6% for δ‐Toc; and 19.6% and 21.8% for total VE. QTL intervals BARCSOYSSR_15_0790–BARCSOYSSR_15_0855 (Qα15_1, Qγ15_1), BARCSOYSSR_15_1113–BARCSOYSSR_15_1159 (Qα15_3, Qδ15_2, QTVE15_4) and BARCSOYSSR_15_1159–BARCSOYSSR_15_1190 (Qα15_4, Qγ15_5, QTVE15_5) were associated with α‐Toc and explained 22.2%, 23.8% and 24.4% of the phenotypic variation in multiple environments. BARCSOYSSR_09_1098–BARCSOYSSR_09_1128 (QTVE9_1) and BARCSOYSSR_15_0887–BARCSOYSSR_15_0935 (QTVE15_2, Qγ15_3) associated with total VE content explained 21.8% and 16.4% of the phenotypic variation in two environments. These QTLs allow for marker‐assisted selection for cultivars with high VE contents.     

Detection and validation of QTLs associated with seed longevity in rice (Oryza sativa L.)

Seed longevity in rice is a major determinant in seed production and germplasm preservation. In this paper, a recombinant inbred line (RIL) population consisting of 172 lines derived from the cross between Xiang743 and ‘Katy’ was used to map quantitative trait loci (QTLs) for seed longevity (SL) after seed storage for 18 and 30 months under ambient conditions. Two putative QTLs, qSL‐2 and qSL‐8, were detected and located on chromosomes 2 and 8, respectively. qSL‐2 is an allele from Xiang743 allele and increases seed longevity. qSL‐8 was a novel QTL from ‘Katy’ allele and increases seed longevity. qSL‐8 explained 15.29% and 17.35% of the phenotypic variance after seed storage for 18 and 30 months, respectively. Furthermore, qSL‐8 was validated in a secondary population developed by self‐pollination of a residual heterozygous line (RHL) selected from the RIL population, which explained 25.93% of the phenotypic contribution. These results provide an opportunity for map‐based cloning of qSL‐8. Furthermore, qSL‐8 may be a target for improving seed longevity by marker‐assisted selection (MAS) in rice.     

Identification of genetic variation in Brassica napus seeds for tocopherol content and composition using near‐infrared spectroscopy technique

Tocopherol is an essential fat‐soluble nutrient for humans. Increasing the tocopherol content in Brassica napus seeds can add value to rapeseed vegetable oil; this has become an important breeding target. However, there is no efficient and non‐destructive method for selecting rapeseed accessions with high tocopherol contents. Here, we report the first near‐infrared reflectance spectroscopy (NIRS)‐based technique for predicting rapeseed tocopherol content. Individual seed tocopherol compositions were estimated from 373 rapeseed genotypes of different origins. This method and chemical methods produced comparable predicted values of the tocopherol constituents in the seeds. Three equations were generated for the prediction of tocopherol content by using a modified partial least squares (MPLS) model. The total tocopherol content for the determination coefficient of cross‐validation (R²_cv) (0.74), determination coefficient (RSQ) (0.76) and one minus the ratio of unexplained variance to total variance (1‐VR) (0.65) values indicates a strong correlation between the calibration and validation sets. Overall, our model confirmed the NIRS method as feasible for predicting tocopherol content in rapeseed and as an efficient screening tool for future breeding programs.     

Genetic control of seed weight and calcium concentration in chickpea seed

Chickpea, Cicer arietinum L., is a staple protein source in many Asian and Middle Eastern countries. Hence, the mineral content of its seed, especially that of calcium, is of nutritional importance. Calcium is transported through plants and to legume pods almost exclusively via the xylem stream, with Ca accretion in developing seeds resulting primarily from diffusion of Ca from the adjoining pod wall. Thus, for seeds of differing surface‐to‐mass ratios, Ca concentration is expected to correlate inversely with seed weight. The relationship between seed weight and Ca concentration in chickpea seeds was studied using a range of germplasm and derivatives from crosses between types differing in seed Ca concentration. Among the cultivars tested, low seed mass was associated with high Ca concentration. However, the study of hybrid progeny indicated that seed Ca content was mainly determined by genetic factors other than grain weight genes. This finding may assist in future breeding of high nutritional quality chickpea 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.     

Selection for contrasting tocopherol content and profile in Ethiopian mustard

Tocopherols are compounds present in oilseeds with major effect on nutritional and technological properties of seed oils. Brassica spp. mainly contain varying concentrations of α‐tocopherol, with high vitamin E value, and γ‐tocopherol, which contributes to oil stability. The objective of this research was to select for contrasting tocopherol content and profile in Ethiopian mustard (Brassica carinata A. Braun). A collection of 211 F_4:5 lines was evaluated for tocopherol content and profile, and divergent selection was conducted till the S_3:4 generation. Selection resulted in two lines with increased tocopherol content (244 and 243 mg/kg seed as average of four environments), one line with reduced tocopherol content (117 mg/kg), two lines with reduced α/γ‐tocopherol ratio (0.22 and 0.28) and two lines with increased α/γ‐tocopherol ratio (3.74 and 3.34). Control line C‐101 averaged 196 mg/kg with α/γ‐tocopherol ratio of 1.10. Reduced α/γ‐tocopherol ratio was controlled by recessive alleles at a single locus. Seed to plant correlation was high (r = 0.92), which suggests the feasibility of selecting at the single‐seed level.     

Identification of quantitative trait loci controlling soybean seed weight in recombinant inbred lines derived from PI 483463 (Glycine soja) × ‘Hutcheson’ (G. max)

The objective of this study was to identify quantitative trait loci (QTLs) controlling 100‐seed weight in soybean using 188 recombinant inbred lines (RIL) derived from a cross of PI 483463 and ‘Hutcheson’. The parents and RILs were grown for 4 years (2010–2013), and mature, dry seeds were used for 100‐seed weight measurement. The variance components of genotype (a), environment (e) and a × e interactions for seed weight were highly significant. The QTL analysis identified 14 QTLs explaining 3.83–12.23% of the total phenotypic variation. One of the QTLs, qSW17‐2, was found to be the stable QTL, being identified in all the environments with high phenotypic variation as compared to the other QTLs. Of the 14 QTLs, 10 QTLs showed colocalization with the seed weight QTLs identified in earlier reports, and four QTLs, qSW5‐1, qSW14‐1, qSW15‐1 and qSW15‐2, found to be the novel QTLs. A two‐dimensional genome scan revealed 11 pairs of epistatic QTLs across 11 chromosomes. The QTLs identified in this study may be useful in genetic improvement of soybean seed weight.     

Evidence of a partial reproductive barrier between wild and cultivated pearl millets (Pennisetum glaucum)

Summary The occurrence of seed malformation in association with reduced thousand grain weight and germination ability has been observed in crosses between cultivated female plants and wild male plants. A survey of 16 cultivated accessions (P. glaucum subsp. glaucum) and 11 wild accessions (P. glaucum subsp. monodii) ranging over the whole species diversity showed this postzygotic incompatibility was general, but its intensity varied greatly with the cultivated female accession used and very little with the wild male parent origin. About 15% of the 123 cultivated x wild crosses observed gave normal seeds. Seed malformation has never been observed in crosses between cultivated accessions and appeared independent of genetic distances between the parents. The reciprocal crosses between wild female plants and cultivated male plants gave normal-looking seeds with good germination but consistently reduced thousand grain weight. Both seed malformation and seed small size are an expression of a genetic imbalance. These slight reproductive barriers seem to have been built during the domestication process.Abbreviation ICRISAT International Crop Research Institute for the Semi-Arid Tropics     

Quantitative dissection of antioxidative bioactive components in cultivated and wild sesame germplasm reveals potentially exploitable wide genetic variability

Sesame (Sesamum indicum L.) is one of the oldest oilseed crops grown in India and worldwide. This oilseed crop has high nutritional value due to the presence of antioxidants such as lignans and tocopherols. Screening of oilseed germplasm for important nutritional attributes is of prime importance in quality breeding programs. In the present study, the content of lignans (sesamin and sesamolin) and tocopherol homologues (α-, γ-, and δ-tocopherol) was determined using reverse phase HPLC (RP-HPLC) in 143 sesame lines collected from diverse agro-ecological zones of India. Exploitable levels of sesamin, sesamolin, γ-, and δ-tocopherol was observed in the studied sesame lines. Sesamum indicum cultivar CO 1, introgressed line MKN 9, and Sesamum malabaricum showed high values for sesamin. Exotic and indigenous accessions of S. indicum (EC 542283 and IC 132176, IC 204681, IC 204773) showed high sesamolin contents. Cultivars, AKT 64, AKT 101, Phule til 1, and Tapi A showed high values for γ- and δ-tocopherol. The average content of sesamin and sesamolin was 0.86 and 0.50 mg g^?1 seed, respectively. The average tocopherol content (292 μg g^?1 seed) found in this study indicates the presence of a high amount of tocopherol in Indian sesame germplasm. Superior genotypes of sesame reported in this study could be utilized in sesame breeding programs for enhancing oil yield and nutritional attributes.     

Detection of loci controlling seed glucosinolate content and their association with Sclerotinia resistance in Brassica napus

A genetic linkage map of Brassica napus constructed from a cross between a low glucosinolate cultivar ‘H5200’ and a high glucosinolate line ‘NingRS‐1’ was used to identify loci associated with seed glucosinolate content and to understand the association between specific glucosinolate components and Sclerotinia resistance. Seed glucosinolate content was assessed by standard High pressure Liquid Chromatogram (HPLC) protocol. Seven components of seed glucosinolate, including four types of aliphatic glucosinolate, two types of indolyl glucosinolates and one aromatic glucosinolate were detected in the seeds. Three quantitative trait loci (QTLs) were identified for seed total glucosinolate content. From three to 15 loci were found to be responsible for different types of glucosinolates, and by comparing the overlapped intervals, eight genomic regions were defined. One of the nine loci associated with aliphatic glucosinolate content was found to be associated with Sclerotinia resistance on the leaf at the seedling stage, and one locus, responsible for 3‐indolyl‐methyl glucosinolate content, was probably linked with Sclerotinia resistance on the stem of the maturing plant. The association between seed glucosinolate content and Sclerotinia resistance is discussed.     

Recurrent selection for increased protein content in yellow mustard (Sinapis alba L.)

Increasing protein content is an important objective in breeding high protein oilseed yellow mustard (Sinapis alba L). The objectives of this research were to increase meal protein content, study population variation during three cycles of selection for increased meal protein content, and quantify the relationships of protein with oil and 1000‐seed weight. Recurrent selection was employed with half‐sib family evaluation in replicated field trials. Meal protein content increased by an average of 1 % per cycle. The correlation between meal protein and seed oil content was negative (r= ‐0.49 to ‐0.58). The population shifts, with selection, reflected successful increase of average meal protein content, and an increased frequency of genotypes with high meal protein content. Furthermore, simultaneous selection for meal protein and seed oil content was possible. The correlation between meal protein content and seed weight was positive (r= 0.29‐0.39) and thus selecting for increased meal protein content posed no risk of decreasing seed weight in this yellow mustard germplasm.     

Comparing mating designs to restore seed production of interspecific hybrids between Trifolium repens (white clover) and Trifolium uniflorum

Interspecific hybrids between Trifolium uniflorum and cultivated white clover (Trifolium repens) have highly useful characteristics for temperate pastoral systems derived from both parent species. However, the early hybrids (F₁ and BC₁) also have unacceptably poor seed production for commerce. This study analysed the basis for the poor seed production and investigated breeding strategies for overcoming the problem. The BC₁F₁ generation produced lower‐than‐expected numbers of heads per plant and seeds per floret. Backcrossing of selected hybrids to white clover corrected these deficiencies and created new variation. Seed numbers were also returned to near target levels by recurrent selection within the BC₁ generation. Thus, it was possible to retain a theoretical average of 25% of T. uniflorum genome and still achieve high seed production per plant. The BC₁F₂ and BC₂F₁ generations produced high seed numbers per plant, along with reasonable variation. Both of these second‐generation hybrid forms have high reproductive potential and should be the focus for the selection of the desired combinations of agronomic and seed production traits.     

红花种子萌发期抗旱性鉴定与评价

为了探索红花耐旱机制及为选育耐旱品种提供理论依据,本文研究了干旱胁迫对红花种质萌发及生长状况的影响。采用聚乙二醇(PEG-6000)模拟干旱胁迫处理,对干旱胁迫下10个红花种质萌发率及幼苗生长指标进行测量,并对各测量指标进行模糊隶属函数分析。结果表明低浓度的PEG对红花种子萌发有一定程度的促进作用；当浓度为15%时,对各指标的抑制作用明显,因此本研究选用15%的PEG浓度鉴定红花种质的抗旱性。在干旱胁迫下红花发芽势、发芽率、下胚轴重、子叶重等降低,胚芽生长受到抑制,干旱对不同种质的根长和根重的影响不一致。通过种子萌发抗旱指数及隶属函数值对红花种质萌发期抗旱性进行综合评价,其中BXY1465为萌发期抗旱性强的材料,BXY512、BXY426、BXY1466以及BXY1859属于萌发期抗旱性中等的材料,BXY1805、BXY2527、BXY2110、BXY750以及WH0086属于萌发期抗性弱的材料。     

The multilocular trait of rapeseed is ideal for high‐yield breeding

The multilocular trait is generally considered as a desirable agricultural trait for the high‐yield breeding of Brassica crops and has attracted considerable attention. The siliques of multilocular plants contain more chambers and seeds than those of bilocular rapeseed varieties and, thus, have a higher yield. Furthermore, multilocular plants are precious germplasm resources for the breeding of Brassica crops. In this article, we provide a summary of researches regarding the multilocular trait in rapeseed, including its natural germplasms, silique morphology, genetic models, and regulatory mechanisms. The study will highlight the potential of multilocular rapeseed for high‐yield breeding.