High regeneration potential in vitro of sunflower (Helianthus annuus L.) lines derived from interspecific hybridization

Successful selection of interspecific hybrid progenies with superior ability to regenerate shoots from apical meristems was performed in sunflower which now allows for the development of lines for improved biotechnological applications. Early generations of interspecific hybrids originating from crosses between the two H. annuus CMS lines ‘HA89’ and ‘Baso’, and 9 wild species were screened for their ability to regenerate in vitro. Evaluation of 36 progenies allowed to identify seven progenies from crosses involving H. mollis, H. giganteus, H. strumosus, and H. decapetalus which showed a significantly higher regeneration potential than the commercial hybrid ‘Albena’ regarding the number of shoots per explant. Among these progenies, 47.2 to 62.4% of explants produced shoots with an average of 2.3 to 3.5 shoots per cultured explant. Regeneration in vitro was significantly determined by the genotype. More than half of the investigated interspecific hybrids performed better than the inbred ‘HA89’ demonstrating that the high regeneration potential available in the wild species can be efficiently transferred to cultivated sunflower. The seven progenies with high regeneration potential in vitro were characterised by agronomic performance in the field. Two of the interspecific hybrids derived from H. strumosus and H. decapetalus not only showed a superior regeneration potential but also proved to be competitive to commercial hybrids with regard to important agronomic traits, e.g. fat content and TGW. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of source germplasm and season on the in vivo haploid induction rate in tropical maize

For in vivo production of doubled haploid (DH) lines in maize, the rate of haploid induction is of crucial importance. Maternal haploid induction depends primarily on the inducer used as a pollinator. However, the source germplasm used as a maternal parent and the environmental conditions for induction may also influence haploid induction and these aspects have not been examined in tropical maize so far. The objectives of our study were to (i) monitor the variation for haploid induction rate (HIR) among diverse source germplasm in tropical maize, (ii) determine the relative importance of general (GCA) and specific (SCA) combining abilities for HIR, and (iii) investigate the influence of summer and winter seasons and genotype × season interactions on this trait. Ten inbreds were mated in a half diallel design. The resulting 45 F₁ single crosses were pollinated with the haploid inducer hybrid RWS × UH400 during the summer 2008 and winter 2009 seasons in a lowland tropical environment in Mexico. HIR of the single crosses averaged over seasons ranged from 2.90 to 9.66% with an overall mean of 6.74%. Mean HIR was significantly (P < 0.01) higher during the winter (7.37%) than summer season (6.11%). Significant (P < 0.01) variation was observed due to GCA effects of parental inbreds of single crosses but not for SCA, GCA × season and SCA × season interactions. Our study underpins that a higher HIR in tropical maize can be obtained by selecting appropriate source germplasm and undertaking pollination under favorable environmental conditions.     

Development of locally-adapted faba bean cultivars for organic conditions in Germany through a participatory breeding approach

Organic farming requires cultivars that are specifically adapted to this low input cropping system. Hence, organic farmers and scientists joined in a participatory breeding approach to develop region-specific genotypes of spring faba bean for organic conditions in Germany. A set of 49 genotypes with contrasting degrees of heterozygosity and heterogeneity was used in field trials across five locations in Germany during 3 years 2004, 2005 and 2006. The material involved 18 inbred lines, their 18 polycross progenies, one blend of inbred lines, one blend of polycross progenies, one blend of hybrids and ten checks. Inbred lines are uniform, thus giving the option to be specifically adapted; whereas the polycross progenies and synthetics (Syn-1; predicted from the inbred lines and polycross progenies performance) are partly heterogeneous and heterozygous, thus giving the option to evolve. Agronomic performance was assessed and a “personal appreciation” score of the material was assigned to each genotype by each partner. This personal appreciation was strongly influenced by biotic and abiotic constraints faced by the crop in each location and by the expected grain yield of the genotypes. Uniformity was apparently appreciated by organic farmers. In all locations, the highest yielding inbred line yielded slightly better than the predicted highest yielding synthetic. However, this slight disadvantage of the synthetic is very likely to disappear if the synthetic (Syn-1) is propagated during successive generations.     

Effect of hardening on frost tolerance and fatty acid composition of leaves and stems of a set of faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.) genotypes

Frost tolerance is a main component of winter-hardiness and improving it would promote faba bean (Vicia faba L.) cropping in cool-temperate regions. In many species, leaf fatty acid composition was found to be related to frost tolerance. The objective of this study was to determine, in a representative sample of genotypes, the effect of hardening on leaf and stem (1) frost tolerance and (2) fatty acid composition, and to seek correlations between them. First leaf, second leaf and stem of 31 faba bean genotypes were analyzed after hardening and without hardening. High frost tolerance of known winter genotypes and several experimental lines was shown. Hardening had a significant, positive effect on frost tolerance of all three organs. Stems were on average more frost tolerant than leaves. Hardening induced significant changes in the fatty acid composition: oleic acid decreased significantly in leaves by 3.24% and in stems by 1.77%, whereas linolenic acid increased in leaves by 6.28% and in stems by 9.06%. In stems, correlations between frost tolerance and fatty acid composition were not significant. Correlation coefficients strongly indicated that non-hardened oleic acid content, changes in oleic acid and in linoleic plus linolenic acid content in leaves partly explained their frost tolerance; 0.347 (P < 0.1) < |r| < 0.543 (P < 0.01). The results corroborate the importance of using genetic differences in the fatty acid metabolism in breeding grain legumes for frost tolerance.     

Construction and characterization of a BAC library for sunflower (Helianthus annuus L.)

A bacterial artificial chromosome (BAC) library was constructed using the sunflower (Helianthus annuus L.) restorer line RHA325, which carries the restorer gene Rf1 and the Pl2-gene conferring resistance to downy mildew. High molecular weight DNA was prepared from nuclei using leaf material from two-week old seedlings. The library was constructed using the HindIII site of pBeloBAC11. The current BAC library comprises 104,736 clones. The insert size of the clones varied between 20 and 270 kb, with an average insert size of 60 kb. The whole 1.9× sunflower BAC library was spotted in duplicate on four high-density filters, each carrying 55,296 clones. The content of organellar DNA, which was estimated by colony hybridisation against the mitochondrial probe coxI and the chloroplast probe rbcL, proved to be less than 0.03 and 0.1%, respectively. BAC pools, allowing PCR-based screening, were made and used to identify positive BAC clones for the markers OP-K13_454, closely linked to the restorer gene Rf1. The PCR-based screening was verified by the results obtained for this marker by colony hybridisation.     

Identification of QTLs for phenolic compounds in oilseed rape (<Emphasis Type="Italic">Brassica napus L.</Emphasis>) by association mapping using SSR markers

Association mapping identifies quantitative trait loci (QTLs) by examining the marker-trait associations that can be attributed to the strength of linkage disequilibrium between markers and functional polymorphisms across a set of diverse germplasm. In this study, association mapping was performed to detect QTL-linked and genome wide SSR markers linked to phenolic compounds of extraction meal in a population of 49 genetically diverse oilseed rape cultivars of dark-seeded, winter-type oilseed rape accessions. Correction for population structure was performed using 559 genome wide SSR markers. Results showed that seed colour is an important contributor to seed meal quality. Totally, 52 SSR markers linked to phenolic compounds were detected, five of them being QTL linked markers. Some of these markers were already mapped on Brassica napus chromosomes that contain known QTL controlling oilseed rape meal quality traits. Our results demonstrate that association mapping is a useful approach to complement and enhance previous QTL information for marker-assisted selection.     

Nitrogen retention and plant uptake on a highly weathered central Amazonian Ferralsol amended with compost and charcoal

Leaching losses of N are a major limitation of crop production on permeable soils and under heavy rainfalls as in the humid tropics. We established a field trial in the central Amazon (near Manaus, Brazil) in order to study the influence of charcoal and compost on the retention of N. Fifteen months after organic‐matter admixing (0–0.1 m soil depth), we added ¹⁵N‐labeled (NH₄)₂SO₄ (27.5 kg N ha^–1 at 10 atom% excess). The tracer was measured in top soil (0–0.1 m) and plant samples taken at two successive sorghum (Sorghum bicolor L. Moench) harvests. The N recovery in biomass was significantly higher when the soil contained compost (14.7% of applied N) in comparison to only mineral‐fertilized plots (5.7%) due to significantly higher crop production during the first growth period. After the second harvest, the retention in soil was significantly higher in the charcoal‐amended plots (15.6%) in comparison to only mineral‐fertilized plots (9.7%) due to higher retention in soil. The total N recovery in soil, crop residues, and grains was significantly (p < 0.05) higher on compost (16.5%), charcoal (18.1%), and charcoal‐plus‐compost treatments (17.4%) in comparison to only mineral‐fertilized plots (10.9%). Organic amendments increased the retention of applied fertilizer N. One process in this retention was found to be the recycling of N taken up by the crop. The relevance of immobilization, reduced N leaching, and gaseous losses as well as other potential processes for increasing N retention should be unraveled in future studies.     

Stereoselectivity of the generation of 3-mercaptohexanal and 3-mercaptohexanol by lipase-catalyzed hydrolysis of 3-acetylthioesters

The enantioselectivity of the generation of 3-mercaptohexanal and 3-mercaptohexanol, two potent sulfur-containing aroma compounds, by lipase-catalyzed hydrolysis of the corresponding 3-acetylthioesters was investigated. The stereochemical course of the kinetic resolutions was followed by capillary gas chromatography using modified cyclodextrins as chiral stationary phases. The enzyme preparations tested varied significantly in terms of activity and enantioselectivity (E). The highest E value (E = 36) was observed for the hydrolysis of 3-acetylthiohexanal catalyzed by lipase B from Candida antarctica resulting in (S)-configured thiol products. Immobilization of the enzyme (E = 85) and the use of tert-butyl alcohol as cosolvent (E = 49) improved the enantioselectivity. Modification of the acyl moiety of the substrate (3-benzoylthiohexanal) had no significant impact. The sulfur-containing compounds investigated possess attractive odor properties, and only one of the enantiomers exhibits the pleasant citrus type note.     

The contribution of<Emphasis Type="Italic"> Phragmites australis</Emphasis> litter to methane (CH<Subscript>4</Subscript>) emission in planted and non-planted fen microcosms

The contribution of Phragmites australis (Cav.) Trin. ex Steud. (common reed) litter as an origin of CH ₄-C was studied in a microcosm experiment. ¹⁴C-labelled, dried and ground P. australis root and shoot litter was buried in waterlogged, planted or non-planted fen microcosms. The evolution of ¹⁴CO ₂ and ¹⁴CH ₄ from the pots was monitored during the 35-day experiment. The ¹⁴C activity in the shoots and roots of the plants, soil, and inundation water was also analysed at the end of the experiment. Up to 40% of the released CH ₄-C originated from the added litter, whereas the rest originated from old soil organic matter. The comparison of planted and non-planted pots suggested that the contribution of recent plant-derived C (i.e. root exudates) to CH ₄ emission was negligible. The proportion of litter-derived CH ₄-C was significantly higher in the planted pots, suggesting that the presence of plants enhanced the formation of CH ₄ from litter. The major part of the initial ¹⁴C activity was recovered from the soil. About 3% was recovered from the inundation water, about 10% was emitted as CO ₂, and only <0.01% as CH ₄. However, these results demonstrated that plant litter and old soil organic matter are the major sources of CH ₄-C in fens during the early growth stage of P. australis.     

Nonvolatiles of commercial lime and grapefruit oils separated by high-speed countercurrent chromatography

The nonvolatile fractions of cold-pressed peel oils of Key and Persian lime as well as grapefruit were separated by high-speed countercurrent chromatography (HS-CCC). In addition to the isolation of the main coumarins, psoralens and polymethoxyflavones, a number of minor constituents were enriched and successfully characterized by GC-MS and HPLC-UV. 5,7,8-Trimethoxycoumarin and the cyclical acetals of oxypeucedanin hydrate with citral were determined as new nonvolatile trace constituents of lime oils and confirmed by NMR spectroscopy. The citral oxypeucedaninyl acetals were found particularly in Key lime oil type A, which as a result of the juice-oil contact, is exposed to acidic conditions during industrial processing. Some of the confirmed minor constituents, such as pabulenol, isooxypeucedanin, and oxypeucedanin methanolate in lime as well as auraptenol in grapefruit, may have been generated by hydrolysis-sensitive precursors during CCC separation or their respective industrial processing techniques.