Gene expression during the ripening of tomato fruit was investigated by cDNA cloning and hybrid-select translation. A cDNA library was prepared from poly(A)-containing mRNA from ripe tomato fruit and sreened by differential hybridization. 146 ripening-related cDNA clones were found. Eleven groups and eight unique clones have been identified so far. The sizes of the cloned cDNA inserts were determined and type-members for seven groups were used in hybrid selection experiments. Six of the seven clones encode translation products corresponding to six ripening related polypeptides detected previously by in vitro translation of total cytoplasmic RNA (14). One cDNA group codes for a Mr 48 000 protein that was identified as polygalacturonase on the basis of immunoprecipitation with specific antiserum raised against tomato polygalacturonase. re]19840918 rv]19850613 ac]19850618.

RAPD analysis was applied to onion (Allium cepa) and otherAllium species in order to assess the degree of polymorphism within the genus and to investigate if this approach was suitable for genetic studies of onion. Seven cultivars ofA. cepa, including shallot, and single cultivars of Japanese bunching onion (A. fistulosum), chive (A. schoenoprasum), leek (A. ampeloprasum), and a wild relative of onion (A. roylei), were evaluated for variability using a set of 20 random 10-mer primers. Seven out of the twenty primers revealed scorable polymorphisms between cultivars ofA. cepa and these will be further evaluated for use in genetic mapping. Wide variations in banding profiles between species were observed with nearly every primer tested. These were assessed for use in systematic studies within the genus. Ninety-one band positions were scored (+/-) for all the cultivars studied. Genetic distances between each of the cultivars were calculated and cluster analysis was used to generate a dendrogram showing phylogenetic relationships between them. The resulting analysis was in broad agreement with previous classifications of the species studied, confirming the validity of the method. However, amongst the species studied, it placedA. roylei as the closest relative ofA. cepa, questioning the current classification of the former species in the section Rhizideum.

DNA barcoding methods originally developed for the identification of plant specimens have been applied to the authentication of herbal drug materials for industrial quality assurance. These methods are intended to be complementary to current morphological and chemical methods of identification. The adoption of these methods by industry will be accelerated by the introduction of DNA-based identification techniques into regulatory standards and monographs. The introduction of DNA methods into the British Pharmacopoeia is described, along with a reference standard for use as a positive control for DNA extraction and polymerase chain reaction (PCR). A general troubleshooting chart is provided to guide the user through the problems that may be encountered during this process. Nevertheless, the nature of the plant materials and the demands of industrial quality control procedures mean that conventional DNA barcoding is not the method of choice for industrial quality control. The design of DNA barcode-targeted quantitative PCR and high resolution melt curve tests is one strategy for developing rapid, robust, and reliable protocols for high-throughput screening of raw materials. The development of authentication tests for wild-harvested L. is used as a case study to exemplify these relatively simple tests. By way of contrast, the application of next-generation sequencing to create a complete profile of all the biological entities in a mixed herbal drug is described and its potential for industrial quality assurance discussed.

For a number of years the lipoproteins of serum have been classified as alpha and beta types according to their electrophoretic mobility. Despite this designation very few studies of their electrophoretic properties have been reported. This is chiefly due to the fact that it is very difficult to determine exactly where the lipoproteins migrate when patterns of whole serum are obtained by the classical Tiselius method. Isolation of the lipoproteins can be achieved by chemical (1, 2) and ultracentrifugal methods (3, 4). However, even if turbidity and solubility problems in the electrophoretic separation of the isolated lipoproteins are overcome, some question remains as to whether the mobilities of these delicate complexes are the same in the isolated state as they are in whole serum. To circumvent these difficulties and to supplement the information obtained from the ultracentrifuge by Gofman, Lindgren and Elliott (3), Green, Lewis and Page (4), and Turner and associates (5) and that obtained from chemical fractionation by Lever and associates (2) and Russ, Eder and Barr (6, 7), the new technique of zone electrophoresis was applied to this problem. This method has the advantage that the electrophoretic components can be isolated directly so that lipid analyses can be carried out. The results obtained in the present study indicate that lipoprotein patterns can be obtained on whole serum and can be correlated in terms of relative electrophoretic mobility with the other proteins of serum. MATERIAL AND METHODSSera were obtained from normal individuals and patients in the fasting state unless otherwise indicated. Analyses were carried out within three days after the removal of the serum except in the case of hypothyroidism where the serum had been stored at 40C for 11 months without the development of significant sediment or lipid material at the surface. The patients with primary biliary cirrhosis were part of a group discussed previously (8). Total lipid, cholesterol and phospholipid analyses were carried out on the same alcohol-ether extract of serum or isolated protein fraction. The methods used were essentially those described in a previous report (8 The method of zone electrophoresis was similar to that described previously (10). Filter paper and potato starch were used as the two types of supporting media in the present study. For the filter paper experiment, three to nine sheets of Whatman 3 MM paper 14 inches in length and 4 inches in width were superimposed on each other.A rectangular segment was cut out of the block of filter paper and the segment and block immersed in buffer solution. The wet paper was then dried to a barely moist state on another sheet of thick filter paper and the blocks placed on a silicone treated glass plate. Serum (2-5 cc.) was then applied to the cut segment of paper and this was replaced in its original position in the block of filter paper. Another glass plate was placed on top of the paper and clamps applied. Silicone grease was spread along the edge of the paper block between th...

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