Ethyl m-digallate from red maple, Acer rubrum L., as the major resistance factor to forest tent caterpillar, Malacosoma disstria Hbn.

An ethanolic extract of red maple (Acer rubrum L.) leaves (RME) applied to trembling aspen (Populus tremuloides Michx.) leaves reduced feeding in choice test assays with forest tent caterpillar larvae (Malacosoma disstria Hbn.) (FTC), whereas a trembling aspen foliage extract, similarly applied, stimulated feeding. Compounds isolated from the RME were gallic acid, methyl gallate, ethyl gallate, m-digallate, ethyl m-digallate, 1-O-galloyl-beta-D-glucose, 1-O-galloyl-alpha-L-rhamnose, kaempferol 3-O-beta-D-glucoside, kaempferol 3-O-beta-D-galactoside, kaempferol 3-O-beta-L-rhamnoside, kaempferol-3-O-rhamnoglucoside, quercetin 3-O-beta-D-glucoside, quercetin 3-O-beta-L-rhamnoside and quercetin 3-O-rhamnoglucoside, (-)-epicatechin. (+)-catechin and ellagic acid. All of the gallates, (-)-epicatechin, and kaempferol 3-O-beta-L-rhamnoside deterred feeding on trembling aspen leaf disks when applied at 0.28 mg/cm2. The two digallates deterred feeding by 90% and were the most effective. HPLC analysis indicated that ethyl m-digallate is present in amounts 10-100 x higher in RME (approximately 2.5-250 mg/g) than any other compound. Thus, ethyl m-digallate appears to be the major compound protecting red maple from feeding by FTC, with a minor contribution from other gallates.

Quercetin glycosides in Psidium guajava L. leaves and determination of a spasmolytic principle.

The traditional herbal remedy from Psidium guajava leaves has been medically proposed in Mexico as effective treatment of acute diarrhea. A methanolic leaf extract was subjected to a bioassay-guided isolation of spasmolytic constituents. Six fractions were separated on a polyvinylpolypyrrolidine (PVPP) column using a water methanol-gradient. The fraction containing flavonols inhibited peristalsis of guinea pig ileum in vitro. A trace of quercetin aglycone together with five glycosides was isolated from this active fraction and identified as quercetin 3-O-alpha-L-arabinoside (guajavarin); quercetin 3-O-beta-D-glucoside (isoquercetin); quercetin 3-O-beta-D-galactoside (hyperin); quercetin 3-O-beta-L-rhamnoside (quercitrin) and quercetin 3-O-gentobioside. Biological activity of each pure compound was studied in the same bioassay. Obtained results suggest that the spasmolytic activity of the Psidium guajava leaf remedy is mainly due to the aglycone quercetin, present in the leaf and in the extract mainly in the form of five flavonols, and whose effect is produced when these products are hydrolyzed by gastrointestinal fluid.

Correlation of phenolic acid content of maize to resistance toSitophilus zeamais, the maize weevil, in CIMMYT'S collections.

The (E)-ferulic acid content of the grain of nine populations of land races of maize derived from CIMMYT's collections was found to be negatively correlated to susceptibility characteristics towards the maize weevilSitophilus zeamais. Correlation coefficients for six susceptibility parameters and (E)-ferulic acid content were significant and ranged from -0.58 to -0.79. A multiple regression analysis by the SAS forward procedure using the primary seed characteristics associated with susceptibility indicated that the ferulic acid content was the only significant factor in explaining variation in at least two susceptibility parameters: the Dobie index and adult preference. In 15 CIMMYT pools, correlations between four susceptibility parameters and (E)-ferulic acid content were also significant (-0.76 to -0.81). The results suggest that phenolic acid content is a leading indicator of grain resistance or susceptibility to insects and may represent a newly identified mechanism of resistance.

An investigation of the intracellular site of anthocyanoplasts using isolated protoplasts and vacuoles.

Microscopic observations made during preparation of protoplasts and vacuoles from red radish seedling hypocotyl (Raphanus sativus L.) show that anthocyanoplasts, the strongly pigmented bodies present in the pigmented cells of the hypodermis, begin as apparently membranous vesicles in the cytoplasm made visible by the deposition and accumulation of anthocyanins, but only rarely appear in the isolated vacuole. Isolation of protoplasts and vacuoles was also achieved from mung bean seedling hypocotyl (Vigna radiata L Wilczek), red cabbage leaf (Brassica oleracea L.) and Prunus x yedoensis Matsum callus. Anthocyanoplasts were usually in the vacuole, although sometimes in the cytoplasm, of the mung bean and cabbage, but were never seen in vacuoles of Prunus callus.

Factors contributing to resistance of exotic maize populations to maize weevil,Sitophilus zeamais.

Factors contributing to resistance of maize to infestation by the corn weevilSitophilus zeamais were investigated in four populations of indigenous and improved maize from Belize. Resistance was related to the antifeedant properties of grain as well as sugar content and mechanical hardness. Grain extracts of all populations of maize significantly reduced insect feeding on treated artificial diets when compared to control diets. Consumption of treated diets was negatively correlated with phenolic content of the grain extract. An analysis by GC-MS indicated that ferulic acid andp-coumaric acid were the principal phenolics present in the extracts, and insect feeding was strongly deterred when pure substances were added to insect diets. Fluorescence associated with ferulic acid and related compounds in grain sections was located in the pericarp and aleurone layer and was especially intense in the most resistant grain variety.

Reversal of Abscisic Acid-Induced Stomatal Closure by trans-Cinnamic and p-Coumaric Acid.

Abscisic acid (ABA)-induced increase in stomatal diffusive resistance (SDR) in excised leaves of bean (Phaseolus vulgaris L. cv Pencil Pod) and maize (Zea mays L. cv Golden Bantam) is inhibited by low concentrations of trans-cinnamic acid (TCA) (1 micromolar) and p-coumaric acid (PCA) (10 micromolar) when given together with ABA (10 micromolar) in the transpiration stream through the cut end of the petiole or leaf blade. A concentration effect is observed both in the ABA action and its reversal by phenolic acids. Leaves having attained a high diffusive resistance in ABA solution recover rapidly when transferred to water. ABA (10 micromolar) induced closure of the stomata in onion, Allium cepa L. and Vicia faba epidermal peels. This is associated with loss of K(+) from guard cells. In the presence of TCA (10 micromolar) and PCA (10 micromolar) K(+) is retained in the guard cells with open stomata. The dark closure of stomata is also inhibited by TCA and PCA. It is suggested that these phenolic acids may inhibit the ABA effect by competing with or acting on some ABA-specific site, probably located on the plasma membrane, regulating flux of K(+) ions. A weak association of ABA with the plasma membrane is envisaged because of the rapid recovery obtained upon transferral of the leaves to water.

Morphology of seedling lentil (Lens culinaris Medic., Fabaceae) as influenced by light intensity and heredity.

Lentil seedlings (Lens culinaris Medic.) were grown out-of-doors, in a shaded greenhouse or in a controlled environment chamber (15 degrees C, 14 hr day under a bank of incandescent and fluorescent lights). Bright light promoted branching of seedlings and inhibited internode elongation. Internodes formed after transfer of plants from high to low or from low to high light intensity were like those grown in continuous high or low light intensity. Branching was enhanced following transfer from high to low light intensity. Transfer from low to high light intensity resulted in stimulation of lateral bud development only if irradiance was very high (full sun). Third generation lines from single seed selections exhibited segregation of internode lengths. F4 hybrids of a cross between a "tall" and a "short" plant showed a similar segregation. Number of leaflets per leaf increased from two at nodes 3 and 4 to four at node 7 in the species of lentils examined (L. culinaris, L. orientalis, L. ervoides, L. nigricans) regardless of light intensity.

Homology between legumin-like polypeptides from cereals and pea.

The presence of legumin-like constituents within the globulin fractions of wheat (Triticum aestivum), rye (Secale cereale) and corn (maize, Zea mays) was demonstrated. Two-dimensional analysis of wheat globulins in the presence and absence of a reducing agent revealed the existence of reducible approximately 60 kDa polypeptides. Western-blot analysis with 125I-labelled antibodies raised against the oat (Avena sativa) 12S globulin holoprotein or its alpha-subunits demonstrated, firstly, the immunological homology between the alpha- and beta-subunits of pea (Pisum sativum) legumin and oat 12S globulin, and secondly, the similar occurrence in wheat of antigenically homologous approximately 20kDa and approximately 40 kDa polypeptides that associate via disulphide linkage to form approximately 60 kDa dimers. Western blotting also showed the presence of disulphide-linked approximately 20 kDa and approximately 40 kDa legumin-like subunits within the globulin fractions of rye and corn.

Elongation of the First Internode of the Pea Seedling during Etiolation : Interaction of Environment and Genotype.

In the present study, 40 lines of peas, Pisum sativum L., from the Weibullsholm collection were examined for elongation response of the first internode to growth in soil, sand, or in darkness in air. The results obtained establish that appearance of the first (lowest) node of the epicotyl at the surface of soil was usual in only a few of the lines tested (L25, L1088, L1570). By contrast, appearance of the first node at the surface of sand as a result of enhancement of first internode extension occurred in 80% of the lines tested. In many lines, extension in sand or soil was more than in darkness in air, a surprising observation given that the known response of the pea epicotyl to pressure is to produce the extension inhibitor, ethylene. Results of two crosses, one of tallxdwarf and one of tallxtall, point to the conclusion that the Le/le locus exercises only partial control over first internode extension and that other, as yet unknown, loci are implicated.

The Fate of l-Phenylalanine Fed to Germinating Pea Seeds, Pisum sativum (L.) var. Alaska, during Imbibition.

Radioactive l-phenylalanine-l-(14)C or -U-(14)C was fed to pea seeds during imbibition. More than 95% was imbibed. Less than 1% of the radioactivity was respired as CO(2). Of the radioactivity taken into the embryos, 80% was still in the cotyledons by 3 days. About half of this was unchanged phenylalanine: 5% free, 10 to 20% in soluble proteins, 1 to 6% in cell wall proteins, and 14% released by mild acid hydrolysis. No other radioactive amino acid was found. About 0.3% of the radioactivity was identified as free caffeic, ferulic, and coumaric acids or their glycosides, and a further 5% was released by mild acid hydrolysis into a phenolic acid fraction. About half of the radioactivity in the cotyledons was lost in the fractionation procedures.About 20% of the radioactivity was found in the shootroot axes to which it had been translocated. Phenylalanine accounted for nearly all the identifiable radioactivity: about 1% free, 1 to 3% in soluble proteins, 1 to 5% in cell wall proteins, and 1% bound to insoluble fractions from which it was released by mild acid hydrolysis. About 1% was released into a phenolic acid fraction. About half of the radioactivity in the shoot-root axes was lost in the fractionation procedures.