QTL analyses of fiber components and crude protein in an annual × perennial ryegrass interspecific hybrid population

Annual (Lolium multiflorum Lam.) and perennial (Lolium perenne L.) ryegrasses are two important forage and turfgrass species. Improving the digestibility of forage by decreasing fiber content is a major goal in forage crop breeding programs. An annual × perennial ryegrass interspecific hybrid population was used to map quantitative trait loci (QTLs) for fiber components, neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL), and crude protein (CP). Samples were harvested three times in August and September 2003 and August 2004, respectively. Simple interval mapping was used to detect QTLs from both the male and female parental maps previously developed for the population. Fiber components were all correlated positively with each other and were negatively correlated with CP. The largest correlations were between NDF and ADF with r = 0.86, 0.72, and 0.82 for each of the three harvests. All four traits showed intermediate broad-sense heritability values ranging from 0.35 to 0.72. A total of 63 QTLs were detected for the four traits measured over the three harvests from both the female and male maps. Coincident QTLs were detected on linkage groups (LGs) 2, 6, and 7 for NDF, LGs 1, 2, and 7 for ADF, LGs 6 and 7 for ADL, and LG 2 for CP, respectively. Coincident QTLs were also detected on LGs 2, 6, and 7 for NDF and ADF, providing evidence of the genetic basis of the observed high level of phenotypic correlation. The QTLs on LGs 2, 6, and possibly 7 for fiber components were co-located on the same LG as several lignin biosynthetic genes from perennial ryegrass.     

Differences in rate of ruminal hydrogenation of C18 fatty acids in clover and ryegrass

Biohydrogenation of C18 fatty acids in the rumen of cows, from polyunsaturated and monounsaturated to saturated fatty acids, is lower on clover than on grass-based diets, which might result in increased levels of polyunsaturated fatty acids in the milk from clover-based diets affecting its nutritional properties. The effect of forage type on ruminal hydrogenation was investigated by in vitro incubation of feed samples in rumen fluid. Silages of red clover, white clover and perennial ryegrass harvested in spring growth and in third regrowth were used, resulting in six silages. Fatty acid content was analysed after 0, 2, 4, 6, 8 and 24 h of incubation to study the rate of hydrogenation of unsaturated C18 fatty acids. A dynamic mechanistic model was constructed and used to estimate the rate constants (k, h) of the hydrogenation assuming mass action-driven fluxes between the following pools of C18 fatty acids: C18:3 (linolenic acid), C18:2 (linoleic acid), C18:1 (mainly vaccenic acid) and C18:0 (stearic acid) as the end point. For k_C18:1,C18:2 the estimated rate constants were 0.0685 (red clover), 0.0706 (white clover) and 0.0868 (ryegrass), and for k_C18:1,C18:3 it was 0.0805 (red clover), 0.0765 (white clover) and 0.1022 (ryegrass). Type of forage had a significant effect on k_C18:1,C18:2 (P < 0.05) and a tendency to effect k_C18:1,C18:3 (P < 0.10), whereas growth had no effect on k_C18:1,C18:2 or k_C18:1,C18:3 (P > 0.10). Neither forage nor growth significantly affected k_C18:0,C18:1, which was estimated to be 0.0504. Similar, but slightly higher, results were observed when calculating the rate of disappearance for linolenic and linoleic acid. This effect persists regardless of the harvest time and may be because of the presence of plant secondary metabolites that are able to inhibit lipolysis, which is required before hydrogenation of polyunsaturated fatty acids can begin.     

Mapping of QTLs controlling content of fatty acid composition in rapeseed (Brassica napus)

The improvement of fatty acid composition is one of the major goals of breeding in rapeseed (Brassica napus). The aim of this study was to provide more information on the genetic determination of fatty acid composition by investigating quantitative trait loci (QTLs). The study was based on two-year of field trials (in 2006 and 2007) with a population of recombinant inbred lines (RILs), which originated from a cross between GH06 and P174. The level of erucic acid (C22:1) was significantly negatively correlated with those of palmitic acid (C16:0), oleic acid (C18:1), linoleic acid (C18:2), linolenic acid (C18:3), and eicosenoic acid (C20:1) in both years. A total of 40 QTLs for six fatty acids were detected and most of them were clustered on linkage groups N8, N9, and N13. These results suggested strongly that there were significant correlations between the levels of fatty acid components and would be useful for the future improvement of breeding programs focused on fatty acids in rapeseed.     

Testing water-soluble carbohydrate QTL effects in perennial ryegrass (Lolium perenne L.) by marker selection

Water-soluble carbohydrates (WSC) are an important factor determining the nutritional value of grass forage and development of genetic markers for selection of WSC traits in perennial ryegrass would benefit future breeding programmes. Quantitative trait loci (QTLs) for WSC have been published for an F₂ ryegrass mapping family. Markers showing significant associations with these QTLs were used to design narrow-based populations with homozygosity for target QTLs. Founders were selected from within the mapping family. The divergent populations produced were analysed for WSC content in the glasshouse and the field. There was evidence of complex interactions between WSC content and other factors and traits, including the scale of assessment, time/degree of sward establishment and other forage quality parameters. Differences between the divergent pairs of the various populations were small. However, differences observed between the founder selection groups were maintained and the roles of the QTL regions in regulating forage WSC content were confirmed. In general, the individual divergent populations exploited only a limited extent of the large phenotypic variation available within the mapping family. However, this study sets the scene for exploring the opportunities for marker-assisted breeding strategies for complex traits in obligate out-breeding species, and the challenges of doing this are discussed.     

Seed and agronomic QTL in low linolenic acid, lipoxygenase-free soybean (Glycine max (L.) Merrill) germplasm.

Linolenic acid and seed lipoxygenases are associated with off flavours in soybean products. F5 recombinant inbred lines (RILs) from a cross between a low linolenic acid line (RG10) and a seed lipoxygenase-free line (OX948) were genotyped for simple sequence repeats (SSR), random amplified polymorphic DNA (RAPD), sequence-tagged sites (STS), and cleaved amplified polymorphic sequence (CAPS) markers and evaluated for seed and agronomic traits at 3 Ontario locations in 2 years. One hundred twenty markers covering 1247.5 cM were mapped to 18 linkage groups (LGs) in the soybean composite genetic map. Seed lipoxygenases L-1 and L-2 mapped as single major genes to the same location on LG G13-F. L-3 mapped to LG G11-E. This is the first report of a map position for L-3. A major quantitative trait locus (QTL) associated with reduced linolenic acid content was identified on LG G3-B2. QTLs for 12 additional seed and agronomic traits were detected. Linolenic acid content, linoleic acid content, yield, seed mass, protein content, and plant height QTL were present in at least 4 of 6 environments. Three to 8 QTLs per trait were detected that accounted for up to 78% of total variation. Linolenic acid and lipoxygenase loci did not overlap yield QTL, suggesting that it should be possible to develop high-yielding lines resistant to oxidative degradation by marker-assisted selection (MAS).     

Serum fatty acid analysis and digestibility study in the Vancouver Island marmot (Marmota vancouverensis) fed a captive diet supplemented with α‐linolenic acid

Mammals that hibernate must rely on endogenous lipid reserves to survive over winter. This study was conducted to compare the difference in serum fatty acid composition, dietary intake, and apparent digestibility in the Vancouver Island marmot (N = 6) fed the Metro Zoo lagomorph diet supplemented with α‐linolenic acid [C(18:3) n‐3 (α‐LA)]. The experiment was designed as a 3 × 3 Latin square with three 17‐day collection periods. The test diets contained 12.16, 14.85, and 17.05% α‐LA as a percentage of fatty acids in the diet supplied through the addition of flaxseed oil (～53% α‐LA). Across treatments, dry matter intake (g/d), dry matter digestibility, apparent fat digestibility, and apparent neutral detergent fiber digestibility did not differ significantly (P > 0.05). There were no significant differences in serum α‐LA concentration between the three levels of α‐LA supplementation. However, this supplementation did elevate serum α‐LA, eicosapentaenoic acid [C20:5 (n‐3)], and docosahexaenoic acid [C22:6 (n‐3)] levels compared with feeding the basal zoo lagomorph diet (P < 0.05). Thus, supplementation of the basal zoo lagomorph diet with α‐LA elevated the serum levels of essential fatty acids in the Vancouver Island marmot. Zoo Biol 20:251–259, 2001. © 2001 Wiley‐Liss, Inc.     

QTL analysis and comparative genomics of herbage quality traits in perennial ryegrass (Lolium perenne L.)

Genetic control of herbage quality variation was assessed through the use of the molecular marker-based reference genetic map of perennial ryegrass (Lolium perenne L.). The restriction fragment length polymorphism (RFLP), amplified fragment length polymorphism (AFLP) and genomic DNA-derived simple sequence repeat-based (SSR) framework marker set was enhanced, with RFLP loci corresponding to genes for key enzymes involved in lignin biosynthesis and fructan metabolism. Quality traits such as crude protein (CP) content, estimated in vivo dry matter digestibility (IVVDMD), neutral detergent fibre content (NDF), estimated metabolisable energy (EstME) and water soluble carbohydrate (WSC) content were measured by near infrared reflectance spectroscopy (NIRS) analysis of herbage harvests. Quantitative trait locus (QTL) analysis was performed using single-marker regression, simple interval mapping and composite interval mapping approaches, detecting a total of 42 QTLs from six different sampling experiments varying by developmental stage (anthesis or vegetative growth), location or year. Coincident QTLs were detected on linkage groups (LGs) 3, 5 and 7. The region on LG3 was associated with variation for all measured traits across various experimental datasets. The region on LG7 was associated with variation for all traits except CP, and is located in the vicinity of the lignin biosynthesis gene loci xlpomt1 (caffeic acid-O-methyltransferase), xlpccr1 (cinnamoyl CoA-reductase) and xlpssrcad 2.1 (cinnamyl alcohol dehydrogenase). Comparative genomics analysis of these gene classes with wheat (Triticum aestivum L.) provides evidence for conservation of gene order over evolutionary time and the basis for cross-specific genetic information transfer. The identification of co-location between QTLs and functionally associated genetic markers is critical for the implementation of marker-assisted selection programs and for linkage disequilibrium studies, which will enable future improvement strategies for perennial ryegrass.     

Comparative analysis of multiple disease resistance in ryegrass and cereal crops

Ryegrass (Lolium spp.) is among the most important forage crops in Europe and Australia and is also a popular turfgrass in North America. Previous genetic analysis based on a three-generation interspecific (L. perenne x L. multiflorum) ryegrass population identified four quantitative trait loci (QTLs) for resistance to gray leaf spot (Magneporthe grisea) and four QTLs for resistance to crown rust (Puccinia coronata). The current analysis based on the same mapping population detected seven QTLs for resistance to leaf spot (Bipolaris sorokiniana) and one QTL for resistance to stem rust (Puccinia graminis) in ryegrass for the first time. Three QTLs for leaf spot resistance on linkage groups (LGs) 2 and 4 were in regions of conserved synteny to the positions of resistance to net blotch (Drechslera teres) in barley (Hordeum vulgare). One ryegrass genomic region spanning 19 cM on LG 4, which contained three QTLs for resistance to leaf spot, gray leaf spot, and stem rust, had a syntenic relationship with a segment of rice chromosome 3, which contained QTLs for resistance to multiple diseases. However, at the genome-wide comparison based on 72 common RFLP markers between ryegrass and cereals, coincidence of QTLs for disease resistance to similar fungal pathogens was not statistically significant.     

Genetic Mapping of QTLs Controlling Fatty Acids Provided Insights into the Genetic Control of Fatty Acid Synthesis Pathway in Peanut (Arachis hypogaea L.)

Peanut, a high-oil crop with about 50% oil content, is either crushed for oil or used as edible products. Fatty acid composition determines the oil quality which has high relevance to consumer health, flavor, and shelf life of commercial products. In addition to the major fatty acids, oleic acid (C18:1) and linoleic acid (C18:2) accounting for about 80% of peanut oil, the six other fatty acids namely palmitic acid (C16:0), stearic acid (C18:0), arachidic acid (C20:0), gadoleic acid (C20:1), behenic acid (C22:0), and lignoceric acid (C24:0) are accounted for the rest 20%. To determine the genetic basis and to improve further understanding on effect of FAD2 genes on these fatty acids, two recombinant inbred line (RIL) populations namely S-population (high oleic line ‘SunOleic 97R’ × low oleic line ‘NC94022’) and T-population (normal oleic line ‘Tifrunner’ × low oleic line ‘GT-C20’) were developed. Genetic maps with 206 and 378 marker loci for the S- and the T-population, respectively were used for quantitative trait locus (QTL) analysis. As a result, a total of 164 main-effect (M-QTLs) and 27 epistatic (E-QTLs) QTLs associated with the minor fatty acids were identified with 0.16% to 40.56% phenotypic variation explained (PVE). Thirty four major QTLs (>10% of PVE) mapped on five linkage groups and 28 clusters containing more than three QTLs were also identified. These results suggest that the major QTLs with large additive effects would play an important role in controlling composition of these minor fatty acids in addition to the oleic and linoleic acids in peanut oil. The interrelationship among these fatty acids should be considered while breeding for improved peanut genotypes with good oil quality and desired fatty acid composition.     

Mapping minor QTL for increased stearic acid content in sunflower seed oil

Increased stearic acid (C18:0) content in the seed oil of sunflower would improve the oil quality for some edible uses. The sunflower line CAS-20 (C18:0 genotype Es1Es1es2es2), developed from the high C18:0 mutant line CAS-3 (C18:0 genotype es1es1es2es2; 25% C18:0), shows increased C18:0 levels in its seed oil (8.6%). The objective of this research was to map quantitative trait loci (QTL) conferring increased C18:0 content in CAS-20 in an F₂ mapping population developed from crosses between HA-89 (wild type Es1Es1Es2Es2; low C18:0) and CAS-20, which segregates independently of the macromutation Es1 controlling high C18:0 content in CAS-3. Seed oil fatty acid composition was measured in the F₂ population by gas-liquid chromatography. A genetic linkage map of 17 linkage groups (LGs) comprising 80 RFLP and 19 SSR marker loci from this population was used to identify QTL controlling fatty acid composition. Three QTL affecting C18:0 content were identified on LG3, LG11, and LG13, with all alleles for increased C18:0 content inherited from CAS-20. In total, these QTL explained 43.6% of the C18:0 phenotypic variation. Additionally, four candidate genes (two stearate desaturase genes, SAD6 and SAD17, and a FatA and a FatB thioesterase gene) were placed on the QTL map. On the basis of positional information, QTL on LG11 was suggested to be a SAD6 locus. The results presented show that increased C18:0 content in sunflower seed oil is not a simple trait, and the markers flanking these QTL constitute a powerful tool for plant breeding programs.     

Development of an AFLP-based linkage map and localization of QTLs for seed fatty acid content in condiment mustard (Brassica juncea).

A genetic linkage map of Brassica juncea based on AFLP and RAPD markers was constructed using 131 F1-derived doubled-haploid (DH) plants from a cross between two mustard lines. The map included 273 markers (264 AFLP, 9 RAPD) arranged on 18 linkage groups, and covered a total genetic distance of 1641 cM; 18.3% of the AFLP markers showed a segregation distortion (P < 0.01). The markers with biased segregation were clustered on seven linkage groups. QTLs for oil contents, palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2), linolenic acid (18:3), eicosenoic acid (20:1), and erucic acid (22:1), were mapped on the AFLP linkage map. Correlation studies among fatty acids in the DH population and the localization of QTLs involved in their control indicated that a major gene located on linkage group (LG) 2 controlled the elongation step of erucic acid.     

Multivariate approach to evaluating the fatty acid composition of seed oil in a doubled haploid population of winter oilseed rape (Brassica napus L.)

The fatty acid composition of oil of the zero erucic acid commercial Brassica napus L. is typical for this species. It is rich in oleic acid and contains moderate levels of linoleic and linolenic acid. For human nutrition, it is advantageous primarily to obtain the highest possible content of oleic acid and to maintain the 2:1 ratio of linoleic to linolenic acid, while preserving the average total content of saturated acids. Uni- and multivariate analyses of variance were used for evaluation of doubled haploid lines of winter oilseed rape in respect of five fatty acids: palmitic (C16:0), stearic (C18:0) oleic (C18:1), linoleic (C18:2) and linolenic (C18:3). Some proposals of studying doubled haploid (DH) lines with the use of canonical transformation were also given. In MANOVA, the five original variables (individual fatty acids) were replaced by three 'new' variables (combinations of these acids) and used to evaluate DH lines with respect to the requirements concerning the nutritional role of fatty acids. The first variable was the total content of the saturated acids (C16:0 + C18:0), the second (unchanging) was the content of the monounsaturated acid C18:1, and the third was the difference between polyunsaturated acids, i.e. between linoleic acid, and the doubled content of linolenic acid (C18:2 - 2 x C18:3).     

Discovery and genetic mapping of single nucleotide polymorphisms in candidate genes for pathogen defence response in perennial ryegrass (<Emphasis Type="Italic">Lolium perenne</Emphasis> L.)

Susceptibility to foliar pathogens commonly causes significant reductions in productivity of the important temperate forage perennial ryegrass. Breeding for durable disease resistance involves not only the deployment of major genes but also the additive effects of minor genes. An approach based on in vitro single nucleotide polymorphism (SNP) discovery in candidate defence response (DR) genes has been used to develop potential diagnostic genetic markers. SNPs were predicted, validated and mapped for representatives of the pathogenesis-related (PR) protein-encoding and reactive oxygen species (ROS)-generating gene classes. The F(1)(NA(6) x AU(6)) two-way pseudo-test cross population was used for SNP genetic mapping and detection of quantitative trait loci (QTLs) in response to a crown rust field infection. Novel resistance QTLs were coincident with mapped DR gene SNPs. QTLs on LG3 and LG7 also coincided with both herbage quality QTLs and candidate genes for lignin biosynthesis. Multiple DR gene SNP loci additionally co-located with QTLs for grey leaf spot, bacterial wilt and crown rust resistance from other published studies. Further functional validation of DR gene SNP loci using methods such as fine-mapping and association genetics will improve the efficiency of parental selection based on superior allele content.     

A novel quantitative trait locus on chromosome A9 controlling oleic acid content in Brassica napus

One of the most important goals in the breeding of oilseed crops, including Brassica napus, is to improve the quality of edible vegetable oil, which is mainly determined by the seed fatty acid composition, particularly the C18:1 content. Previous studies have indicated that the C18:1 content is a polygenic trait, and no stable quantitative trait loci (QTLs) except for FAD2 have been reported. By performing a GWAS using 375 low erucic acid B. napus accessions genotyped with the Brassica 60K SNP array and constructing a high‐density SNP‐based genetic map of a 150 DH population, we identified a novel QTL on the A9 chromosome. The novel locus could explain 11.25%, 5.72% and 6.29% of phenotypic variation during three consecutive seasons and increased the C18:1 content by approximately 3%–5%. By fine mapping and gene expression analysis, we found three potential candidate genes and verified the fatty acids in a homologous gene mutant of Arabidopsis. A metal ion‐binding protein was found to be the most likely candidate gene in the region. Thus, the C18:1 content can be further increased to about 80% with this novel locus together with FAD2 mutant allele without compromise of agronomic performance. A closely linked marker, BnA129, for this novel QTL (OLEA9) was developed so that we can effectively identify materials with high C18:1 content at an early growth stage by marker‐assisted selection. Our results may also provide new insight for understanding the complex genetic mechanism of fatty acid metabolism.     

Quantitative trait locus analysis of unsaturated fatty acids in a recombinant inbred population of soybean

Soybean [Glycine max (L.) Merr.] is an important oilseed crop which produces about 30 % of the world’s edible vegetable oil. The quality of soybean oil is determined by its fatty acid composition. Soybean oil high in oleic and low in linolenic fatty acids is desirable for human consumption and other uses. The objectives of this study were to identify quantitative trait loci (QTLs) for unsaturated fatty acids and to evaluate the genetic effects of single QTL and QTL combinations in soybean. A population of recombinant inbred lines derived from the cross of SD02-4-59 × A02-381100 was evaluated for fatty acid content in seven environments. In total, 516 polymorphic single nucleotide polymorphism markers, 477 polymorphic simple sequence repeat markers and three GmFAD3 genes were used to genotype the mapping population. By using the composite interval mapping and/or the interval mapping method, a total of 15 QTLs for the three unsaturated fatty acids were detected in more than two environments. Two QTLs for oleic acid on linkage groups G [chromosome (Chr) 18] (qOLE-G) and J (Chr 16) (qOLE-J), three QTLs for linoleic acid on linkage groups A1 (Chr 5) (qLLE-A1) and G (Chr 18) (qLLE-G-1 and qLLE-G-2), and five QTLs for linolenic acid on linkage groups C2 (Chr 6), D1a (Chr 1), D1b (Chr 2), F (Chr 13) and G (Chr 18) were consistently detected in at least three individual environments and the average data over all environments. Significant QTL × QTL interactions were not detected. However, significant QTL × environment interactions were detected for all the QTLs which were repeatedly detected. Some QTLs reported previously were confirmed, and seven new QTLs (two for oleic acid, two for linoleic acid and three for linolenic acid) were identified in this study. Comparisons of two-locus and three-locus combinations indicated that cumulative effects of QTLs were significant for all the three unsaturated fatty acids. QTL pyramiding by molecular marker-assisted breeding would be an appropriate strategy for the improvement of unsaturated fatty acids in soybean.     

Identifying genetic components controlling fertility in the outcrossing grass species perennial ryegrass (Lolium perenne) by quantitative trait loci analysis and comparative genetics

Mutational load and resource allocation factors and their effects on limiting seed set were investigated in ryegrass by comparative mapping genomics and quantitative trait loci (QTL) analysis in two perennial ryegrass (Lolium perenne) mapping families sharing common genetic markers. Quantitative trait loci for seed-set were identified on chromosome (LG) 7 in both families and on LG4 of the F2/WSC family. On LG7, seed-set and heading date QTLs colocalized in both families and cannot be unequivocally resolved. Comparative genomics suggests that the LG7 region is syntenous to a region of rice LG6 which contains both fertility (S5(n)) and heading date (Hd1, Hd3a) candidate genes. The LG4 region is syntenous to a region of rice LG3 which contains a fertility (S33) candidate gene. QTL maxima for seed-set and heading date on LG4 in the F2/WSC family are separated by c. 8 cm, indicating distinct genetic control. Low seed set is under the control of recessive genes at both LG4 and LG7 locations. The identification of QTLs associated with seed set, a major component of seed yield in perennial ryegrass, indicates that mutational load associated with these genomic regions can be mitigated through marker-assisted selection.     

RISING WATER TEMPERATURES ALTER LIPID DYNAMICS AND REDUCE N‐3 ESSENTIAL FATTY ACID CONCENTRATIONS IN SCENEDESMUS OBLIQUUS (CHLOROPHYTA)1

The biosynthesis of nutritionally important polyunsaturated fatty acids (PUFAs) in phytoplankton is influenced by environmental temperature. We investigated the potential of climate warming to alter lipid dynamics of Scenedesmus obliquus (Turpin) Kütz. by comparing lipid and fatty acid (FA) profiles as well as FA metabolism (using [1‐¹⁴C] acetate) at 20°C and 28°C. We documented an overall decline (53%–37%) in the proportion of n‐3 PUFA (in particular, of α‐linolenic acid [ALA; 18:3n‐3]), and a concomitant increase in saturated fatty acids (SAFAs) in total lipids (TLs) at 28°C, consistent with enhanced incorporation of radioactivity from [1‐¹⁴C] acetate into total 16:0, 18:1, and decreased incorporation into 18:2 and 18:3 FA (from 36% to 22% of the total) at 28°C. Glycerophospholipids were also affected by warming; ALA and stearidonic acids (SDAs; 18:4n‐3) both decreased (by 13% and 15%, respectively) in phosphatidylcholine (PC) and (by 24% and 20%, respectively) in phosphatidylethanolamine (PE). The characteristic FA in phosphatidylglycerol (PG; 16:1n‐13t) increased (by 22%) at 28°C. The activities of desaturases, which add double bonds to FA moieties, comprised the major suite of reactions affected by the temperature increase in TL and polar lipid (PL) classes. Climate modelers predict an increase in the number of extreme heat days in summer at temperate latitudes, with parallel projected increases in water temperatures of shallow water bodies. Our results suggest that the overall decrease in the essential n‐3 FA ALA in S. obliquus at higher water temperatures may lower food quality for higher tropic levels, adding another climate‐warming stress.     

Identification of QTLs for morphological traits influencing waterlogging tolerance in perennial ryegrass (Lolium perenne L.)

Perennial ryegrass is a globally cultivated obligate outbreeding diploid species (2n = 2x = 14) which is subjected to periods of waterlogging stress due to flood irrigation during winter and the lead-up to summer. Reduction of oxygen supply to root systems due to waterlogging produces consequent deleterious effects on plant performance. Framework genetic maps for a large-scale genetic mapping family [F?(NA(x) × AU?)] were constructed containing 91 simple sequence repeat and 24 single nucleotide polymorphism genetic markers. Genetic trait dissection using both control and waterlogging treatments was performed in the glasshouse, a total of 143 maximally recombinant genotypes being selected from the overall sib-ship and replicated threefold in the trial. Analysis was performed for nine quantitative morphological traits measured 8 weeks after stress treatments were applied. A total of 37 quantitative trait loci (QTLs) were identified; 19 on the NA(x) parental genetic map, and 18 on the AU? parental genetic map. Regions of particular interest were identified on linkage groups (LGs) 4 and 3 of the respective maps, which have been targeted for further analysis by selection of critical recombinants. This first study of genetic control of waterlogging tolerance in ryegrasses has important implications for breeding improvement of abiotic stress adaptation.     

Mapping of QTLs for oil content and fatty acid composition in Indian mustard [Brassica juncea (L.) Czern. and Coss.]

An AFLP linkage map of Brassica juncea (L.) Czern and Coss was constructed using 88 recombinant inbred lines (RILs) from a cross between an Indian cultivar ‘Varuna’ and an accession from Poland ‘BEC-144’. The map included 91 AFLP markers organized on 19 linkage groups covering a total map distance of 1679.1 cM. A total of 14 QTLs were detected for oil content (2 QTLs), erucic acid (2 QTLs), eicosenoic acid (2 QTLs), linolenic acid (3 QTLs), linoleic acid (3 QTLs) and palmitic acid (2 QTLs). A specific genomic region on LG2 was associated with contents of three fatty acids: erucic acid, eicosenoic acid and linoleic acid. Some of the markers showed absolute linkage with the QTLs associated with the levels of linolenic acid, linoleic acid and oil content. These markers may be used for improvement of fatty acid profile of B. juncea.     

Changes in fatty acid composition,cholesterol and fat‐soluble vitamins during development of eggs and larvae in shabbout (Barbus grypus,Heckel 1843)

Changes in fatty acid composition, cholesterol and fat‐soluble vitamins were studied during development (fertilized eggs, yolk‐sac larvae, and after yolk resorption of shabbout, Barbus grypus). Significant differences were found in the monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFA), PUFA/saturated fatty acids (SFA), ∑n‐3 and eicosapentaenoic acid (EPA)/docosahexaenoic acid (DHA) ratios between eggs and larvae (P < 0.05). Significant differences were also observed in the C14:0, C16:1n‐7, C18:1n‐9, C18:3n‐6, C20:0, C20:4n‐6, C24:0, C24:1, C22:6n‐3 fatty acids between eggs and larvae after yolk‐sac resorption (P < 0.05). Vitamin α‐ Tocopherol and retinol content increased during embryogenesis, but changes were insignificant in retinol acetate, δ‐Tocopherol, K1, K2 and cholesterol content between eggs and larvae after yolk resorption (P > 0.05).