TILLING and Ecotilling the Plant Kingdom

The modem crop scientist has a large amount of available nucleotide sequence information to identify genes of potential agronomic importance. Using reverse genetic approaches, specific genes can be disrupted, and hypotheses regarding gene function directly tested in vivo. Although a number of reverse genetic methods have been introduced, many are limited in application because they are organism-specific, expensive, transgenic, or only transiently disrupt gene function. However, traditional mutagenesis using chemical mutagens has been widely used as a forward genetics strategy to create many new crop plant varieties at relatively low cost. Mutagens such as ethyl methanesulphonate （EMS） cause stable point mutations and thus produce an allelic series of truncation and missense changes that can provide a range of phenotypes （Greene et al., 2003）. TILLING （targeting induced local lesions IN genomes） is a high-throughput reverse genetic strategy that combines traditional mutagenesis and SNP discovery methods （Colbert et al., 2001; McCallum et al., 2000）. To identify mutations, target regions of-l.5 kb are amplified with fluorescently labeled gene specific primers. Heteroduplexes are then formed between wild-type and mutant strands, mismatches are cleaved by incubation with a single-strand specific nuclease,     

Using model scenarios to predict and evaluate forest-management impacts on soil base saturation at landscape level

Silviculture, forest conversion and technical tools of ecosystem management, such as forest liming, display their effects at the landscape level. Therefore their planning and control should take place at the same scale. The primary objective of this work was to assess soil chemical properties and their changes in relationship to ecosystem management, especially forest conversion and forest liming. We calculated scenario models, based on regression analysis, which allow such an examination in the context of understanding landscape processes which can be expected to operate in the sampling region. Stepwise multiple linear regression analysis was used to predict soil chemical attributes (base saturation, pH, C/N content and stock, exchangeable stocks of Ca and Mg) as indicators of site stability or off-site effects of forest ecosystems such as effects on clean drinking water from forested watersheds. Because of space limitations, in this paper only the modelling results of base saturation are presented. Base saturation was used as an integrative example for other soil chemical properties. The transformation of measurements to the regional scale, i.e., the regionalization, was calculated for the forested parts of two test regions in the Black Forest on the basis of measured chemical properties of 90–150 soil profiles per test region. The models have a spatial resolution of 50×50 m, which is a spatial scale relevant for forestry practice and forest management. Topographic variables (e.g., elevation, aspect, slope gradient, and slope length), the stratigraphic classification of the geologic substrate, stand characteristics from forest inventory data, and finally technical information about forest liming were the auxiliary variables (secondary site properties) that provided indirect information about base saturation and were available for the whole forested area of the test regions. Base saturation could be predicted with an accuracy of ~50–70% (in terms of the multiple R²) by using these properties as predictor variables in multiple linear regression analyses. The explained proportion of variance was unexpectedly high considering the high geomorphological heterogeneity of the two test regions. Based upon the regionalization models, it was possible to establish scenarios showing the landscape-related effects on base saturation that may be achieved by forest conversion towards a higher proportion of forests with broad-leaved mixed stands and by forest liming. These scenarios allow the interactions between several influencing factors and management strategies and the impacts on the target variable to be synoptically judged. Thus the presented regionalization models achieve the role of decision support tools for the planning of forest management at the landscape level. They allow an assessment of the environmental effects of forest management strategies in terms of site sustainability or preservation of water resources in forested catchments.     

Differentiation between lutein monoester regioisomers and detection of lutein diesters from marigold flowers (Tagetes erecta L.) and several fruits by liquid chromatography-mass spectrometry.

Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-APCIMS) was employed for the identification of eight lutein monoesters, formed by incomplete enzymatic saponification of lutein diesters of marigold (Tagetes erecta L.) by Candida rugosa lipase. Additionally, the main lutein diesters naturally occurring in marigold oleoresin were chromatographically separated and identified. The LC-MS method allows for characterization of lutein diesters occurring as minor components in several fruits; this was demonstrated by analysis of extracts of cape gooseberry (Physalis peruviana L.), kiwano (Cucumis metuliferus E. Mey. ex Naud.), and pumpkin (Cucurbita pepo L.). The assignment of the regioisomers of lutein monoesters is based on the characteristic fragmentation pattern: the most intense daughter ion generally results from the loss of the substituent (fatty acid or hydroxyl group) bound to the epsilon-ionone ring, yielding an allylic cation. The limit of detection was estimated at 0.5 microg/mL with lutein dimyristate as reference compound. This method provides a useful tool to obtain further insight into the biochemical reactions leading to lutein ester formation in plants.     

Identification and quantification of zeaxanthin esters in plants using liquid chromatography-mass spectrometry

It has been suggested that lutein and zeaxanthin may decrease the risk for age-related macular degeneration. Surprisingly, oleoresins rich in zeaxanthin are not yet available on the market. Several authors have reported enhanced stability of esterified xanthophylls, so plants containing zeaxanthin esters were investigated to establish valuable sources for the production of durable oleoresins. Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [LC-(APCI)MS] was used to unequivocally identify zeaxanthin esters of a standard mixture and in several plant extracts. Zeaxanthin esters were quantified on the basis of their respective molecular masses using zeaxanthin for calibration; total zeaxanthin was determined after saponification of aliquots of the extracts. Thus, dried wolfberries (Lycium barbarum), Chinese lanterns (Physalis alkekengi), orange pepper (Capsicum annuum), and sea buckthorn (Hippophae rhamnoides) proved to be valuable zeaxanthin ester sources. The present LC-MS method allows for an even more detailed analysis of zeaxanthin esters than reported previously.     

Extraktion eines kupferbelasteten Bodens unter Verwendung eines aminosäurehaltigen Reststoffhydrolysats. 2. Elutionsdynamik und Bilanzierung der Aminosäuren nach kontinuierlichem Eintrag in Bodensäulen

Extraction of a copper contaminated soil material by the percolation of an amino acid containing residue hydrolysate. 2. Time course of amino acid elution and input/output balance of amino acids During 16 days an amino acid containing blood meal hydrolysate (amino acid concentration: 188 mMol·L^?1) was percolated through a column packed soil material (soil content per column: 4.1 kg dry weight, four parallels). The copper contaminated material (soil type: Typic Udifluvent, soil texture: sandy loam, loamy sand) was sampled from an area formerly used for cultivation of hop (Humulus lupus). Besides the investigation of the copper liberation the experiments aimed to determine the elution dynamic and input/output balance of amino acids (time span for amino acids balance 14 days). In total 11.7 L of hydrolysate, containing 2.2 Mol of amino acids, were introduced into each column. The mean amino acid output with the column effluent was 1.13 Mol. This corresponds to an elution degree of 51.2%, related to the sum of applicated amino acids, and to a mean substance specific elution degree of 48.4% reflecting the elution of 15 compounds. The substance specific elution ranged from 9.6% (serine) to 75.5% (valine). The highest concentrations of serine and threonine were determined in the effluents after two days, whereas the histidine concentration was highest at the last sampling. The differences in the percolation properties of the amino acids are discussed in terms of important retention and elimination processes (biodegradation, ad-/desorption, intercalation).     

A method for identification and mapping potentially acid sulfate soils in Jutland, Denmark

A method for mapping potentially acid sulfate soils in Jutland, Denmark is described. First the wetlands were outlined, using already existing topographic maps. Then 7900 profiles, evenly distributed over the wetland areas, were examined to point out areas with potentially acid sulfate soils. The method used to identify potentially acid sulfate material was, for carbonate-free samples, a pH-measuring at the sampling time and after the samples were inoculated with soil water extract from an acid sulfate soil and 2, 8 or 16 weeks of incubation. For carbonate-containing samples the pyrite content was compared to the acid-neutralizing capacity. A map was constructed showing the ratio of profiles containing potentially acid sulfate soil samples to total number of profiles examined within natural geographical wetland regions.The investigation showed that 35% of the wetlands in Jutland have a high frequency of potentially acid sulfate soils.     

Element (Ag, Cd, Cu, Pb, Sb, Tl and Zn), element ratio and lead isotope profiles in a sediment affected by a mining operation episode during the late 19th century

Mining operations at Mårsätter in 1877–81 resulted in increased metal loading to a small lake, notably as sulphidic tailings. The event is taken as an opportunity to study the present environmental impact of a historical single major metal release. Lake water and four sediment cores were sampled and analysed for principal and trace elements in solid and aqueous phases as well as general hydrochemical conditions. Chronologies were determined from ²⁰⁶Pb/²⁰⁷Pb ratios and historical records. Ordinary sedimentation after the event has lead to that the tailings are found as a distinct layer at a depth of 18–22 cm in the sediment. The layer is characterized by elevated metal concentrations in the solid and pore water phases, respectively, circum neutral pH and sulphate concentrations below detection. Geochemical modelling indicated a preference for carbonate equilibrium in the waste while sulphides prevailed above it. It is concluded that the growth of the ordinary sediment on top of the waste has lead to a physicochemical barrier that seals of the waste from the overlying sediment. Chemical or physical rupture of the barrier will release the metals to downstream regions. According to the chronologies at least three sources have contributed to the present elevated levels of metals, in additions to the release of tailings. Copper from a historical blast furnace prior to the event at Mårsätter, transport from mineralized parts of the watershed and release of contaminated water from present mining operations maintain elevated levels of notably zinc, silver, cadmium and lead. At present less than 10% of the lead content at the sediment/water interface comes from atmospheric deposition. Increased levels of antimony and thallium were not observed prior to ca 1950.     

TerraRossa: A Mid-Season Specialty Potato with Red Flesh and Skin and Resistance to Common Scab and Golden Cyst Nematode

TerraRossa (POR01PG20–12) is a mid-season specialty potato, released by Oregon State University, and is a product of the Northwest Potato Variety (Tri-State) Development Program. This cultivar is unique among commercially available potato cultivars in that plants produce small- to medium-sized smooth, oblong- to long-shaped tubers with red skin and red flesh. Total tuber yields of TerraRossa are similar to Dark Red Norland and less than Red La Soda. Average tuber size (136 g) is less than both of the comparison cultivars, reflecting inherent differences in tuber size distribution. TerraRossa tubers have total antioxidant levels higher than traditional white fresh varieties and comparable to the All Blue purple potato, known for its high antioxidant levels. Sensory evaluations of TerraRossa tubers indicated that it has good culinary attributes following boiling, baking, and microwaving. Potato chips made from TerraRossa tubers retained their unique red color, which represents a novelty in the chipping industry. TerraRossa could be a good candidate for the organic sector due to its tolerance to common scab (Streptomyces scabies) and tuber late blight (Phytophthora infestans) and its resistance to golden cyst nematode (Globodera rostochiensis). Due to its high yields, high chipping quality, good culinary properties, high antioxidant content and disease resistance, TerraRossa is a good candidate for opening new specialty type markets, adding diversity to the marketplace.