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,     

From wine to pepper: rotundone, an obscure sesquiterpene, is a potent spicy aroma compound

An obscure sesquiterpene, rotundone, has been identified as a hitherto unrecognized important aroma impact compound with a strong spicy, peppercorn aroma. Excellent correlations were observed between the concentration of rotundone and the mean 'black pepper' aroma intensity rated by sensory panels for both grape and wine samples, indicating that rotundone is a major contributor to peppery characters in Shiraz grapes and wine (and to a lesser extent in wine of other varieties). Approximately 80% of a sensory panel were very sensitive to the aroma of rotundone (aroma detection threshold levels of 16 ng/L in red wine and 8 ng/L in water). Above these concentrations, these panelists described the spiked samples as more 'peppery' and 'spicy'. However, approximately 20% of panelists could not detect this compound at the highest concentration tested (4000 ng/L), even in water. Thus, the sensory experiences of two consumers enjoying the same glass of Shiraz wine might be very different. Rotundone was found in much higher amounts in other common herbs and spices, especially black and white peppercorns, where it was present at approximately 10000 times the level found in very 'peppery' wine. Rotundone is the first compound found in black or white peppercorns that has a distinctive peppery aroma. Rotundone has an odor activity value in pepper on the order of 50000-250000 and is, on this criterion, by far the most powerful aroma compound yet found in that most important spice.     

A microsatellite sequence closely linked to the Waxy gene of Oryza sativa

Summary A polymorphic microsatellite locus has been located closely linked to the Waxy gene of rice. Primers were designed to allow detection of the microsatellite by utilising the polymerase chain reaction. In screen of 13 commercial rice varieties, four different alleles were found, demonstrating the potential of this marker in commercial rice breeding for starch quality.Abbreviations RFLP Restriction Fragment Length Polymorphism - PCR Polymerase Chain Reaxtion     

The Share of Southeastern Black Counties in the Southern Rural Renaissance

Some authors have suggested that a leading problem associated with US factory employment for blacks may be that the nonfarm jobs--specifically manufacturing--have tended to go to heavily white counties rather than to the mainly black counties of the Deep South. This study is limited to the 1959-1977 period, the time of the most vigorous nonmetropolitan employment growth. To test the hypothesis that factory jobs have bypassed heavily black counties, a 100% sample of nonmetropolitan counties was used from 13 Southern states for the 2 time periods, 1959-1969 and 1969-1977. Results show that the minority counties were not bypassed by Southern rural factory job growth either in the 1960s or the 1970s, but their gains were not as sizeable as those of more heavily white counties. Both the neoclassical economics of discrimination--with its individualistic "taste for discrimination"--and the stinging critique by Marshall in 1974 that such analysis ignores the important role of institutionalized racism--can "explain the empirical conclusions of this article." Traditional "smokestack-chasing" methods are by no means hopeless. While co-ops and community development corporation strategies can be combined with traditional methods, it is important that foundations, government agencies, anti-poverty programs, and black leadership also focus on projects to competitively attract jobs in the traditional fashion.     

Improved sample preparation to determine acrylamide in difficult matrixes such as chocolate powder, cocoa, and coffee by liquid chromatography tandem mass spectroscopy

An improved sample preparation (extraction and cleanup) is presented that enables the quantification of low levels of acrylamide in difficult matrixes, including soluble chocolate powder, cocoa, coffee, and coffee surrogate. Final analysis is done by isotope-dilution liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) using d3-acrylamide as internal standard. Sample pretreatment essentially encompasses (a) protein precipitation with Carrez I and II solutions, (b) extraction of the analyte into ethyl acetate, and (c) solid-phase extraction on a Multimode cartridge. The stability of acrylamide in final extracts and in certain commercial foods and beverages is also reported. This approach provided good performance in terms of linearity, accuracy and precision. Full validation was conducted in soluble chocolate powder, achieving a decision limit (CCalpha) and detection capability (CCbeta) of 9.2 and 12.5 microg/kg, respectively. The method was extended to the analysis of acrylamide in various foodstuffs such as mashed potatoes, crisp bread, and butter biscuit and cookies. Furthermore, the accuracy of the method is demonstrated by the results obtained in three inter-laboratory proficiency tests.     

Context matters: the landscape matrix determines the population genetic structure of temperate forest herbs across Europe

Landscape Ecology - Plant populations in agricultural landscapes are mostly fragmented and their functional connectivity often depends on seed and pollen dispersal by animals. However, little is...