Interaction between proteins and glucosinolate isothiocyanates and oxazolidinethiones from Brassica napus seed

The proteins from rape seed meal and serum albumin were incubated with the ³⁵S-labelled glucosinolates progoitrin, gluconapin, and glucoalyssin in a variety of reaction conditions. Intact glucosinolates and oxazolidinethiones were found to combine with the proteins to a very small extent, independently of pH; but the isothiocyanates reacted readily with the proteins at pH values higher than 6. Fractionation of the rape seed protein conjugates on Sephadex G200 showed that isothiocyanates particularly reacted with the basic low molecular weight proteins. Changes in UV-spectrum and electrophoretic mobility after reaction with isothiocyanates were also demonstrated.     

1-Cyano-3,4-epithiobutane: A major product of glucosinolate hydrolysis in seeds from certain varieties of Brassica campestris

A new hydrolysis product derived from 3-butenylglucosinolate in seeds of certain strains of Brassica campestris Yellow Sarson is described. The structure, 1-cyano-3,4-epithiobutane is proposed. If the seeds are heated at 115° for 30 min before hydrolysis, 3-butenyl isothiocyanate is the main product.     

β-Galactosidases of Lilium pollen

Lily (Lilium auratum) pollen contains very high levels of β-galactosidase. There are three forms: β-galactosidase I and II differ in M_r, while β-galactosidase III is firmly bound in the pollen wall. The two cytoplasmic forms were separated and partially purified using a combination of chromatography on DEAE-cellulose, Sephadex G-200 and Sepharose 6B. Forms I and II appear to be glycoprotein in nature as shown by binding to Con A-Sepharose. The three enzymes were optimally active near pH 4, and all were inhibited by galactose and galactonolactone. The wall-bound enzyme, β-galactosidase III effectively hydrolysed nitrophenyl β-galactosidase but not lactose, and could not be released from the wall polysaccharide matrix by high salt concentrations or detergents. The total β-galactosidase activity of lily pollen remained constant during in vitro germination. A possible role for this enzyme may be in degradation of stylar arabinogalactans providing a carbon source for pollen tube nutrition.     

6-d-glucopyranosyl fatty acid esters from Brassica napus pollen

6-d-Glucopyranosyl esters of palmitic, oleic, linoleic and linolenic acids were identified in Brassica napus (rape) pollen. These esters are inactive as plant growth promoters in the bean second-internode bioassay.     

Structural investigation of an arabinan from cabbage (Brassica oleracea var. capitata)

An arabinan has been isolated from the hot water-soluble pectic substances of cabbage cell wall material. Methylation analysis involving GC-MS of methylated alditol acetates formed from the methylated arabinan has shown that the parent polysaccharide is highly branched and of the same structural type as other arabinans associated with seed pectins.     

Glucosinolate hydrolytic products given by Sinapis alba, and Brassica napus thioglucosidases

Thioglucosidase prepared from rapeseed (Brassica napus cvs Zephyr and Bronowski) showed one major band in polyacrylamide gel and a high susceptibil     

Polysaccharide component in the stigmatic exudate from Lilium longiflorum

The polysaccharide component of the stigmatic exudate from Lilium longiflorum has the composition, arabinose (26%), rhamnose (6%), galactose (57%) and glucuronic acid (11%). The highly branched polysaccharide bears a striking resemblance to the acidic polysaccharide exudate from Araucaria bidwillii in belonging to the galactan group and in carrying outer chains terminated by arabinofuranose, rhamnopyranose, galactopyranose and glucuronic acid residues. Both polysaccharides contain the sequence O-rhamnopyranosyl-(1→4)-glucopyranosyluronic acid-(1→6)-galactopyranose in some of the outer chains.     

Sugar composition of pollen grain and pollen tube cell walls

The sugar composition of pollen grain and pollen tube cell walls was studied for Camellia japonica, C. sasanqua, C. sinensis, Tulipa gesneriana and Lilium longiflorum. In all species, the main components of pollen grain walls were arabinose, galactose, glucose and uronic acid. On the other hand, the pollen tube walls consisted mostly of glucose. The pollen tube wall of C. japonica was fractionated into hemicellulose, α-cellulose and pectic substance fractions in yields of 61, 19 and 3 %, respectively. The hemicellulose fraction was composed essentially of glucose. The sugar composition of the pollen tube wall was not influenced by the nature of exogenously supplied sugars. Rapid growth of the pollen tube seemed to correlate with the synthesis of hemicellulosic glucan.     

Pectic polysaccharides of cabbage (Brassica oleracea)

Pectic substances extracted from cabbage cell walls with water, at 80°, and (NH₄)₂C₂O₄, at 80°, accounted for 45%(w/w) of the purified cell wall material. Only a small amount of neutral arabinan was isolated. Partial acid hydrolysis and methylation analysis revealed that the major pectic polysaccharide had a rhamnogalacturonan backbone to which a highly branched arabinan was linked, at C-4 of the rhamnose units, mainly through short chains of (1→4)-linked galactopyranose residues. The bulk of the soluble pectic substances had only small amounts of proteins associated with them. After further extraction of the depectinated material with 1M and 4M KOH, to remove the hemicelluloses, the cellulose residue was found to contain a pectic polysaccharide which was solubilized by treatment with cellulase. The general structural features of the pectic polymers are discussed in the light of these results.     

Wax synthesis in Brassica oleracea as modified by trichloroacetic acid and glossy mutations

Two different mutations in Brassica oleracea, gl₅ and gl₄ have been re-investigated using acetate-1-¹⁴C labelling in an attempt to define more closely the nature of the genetic blocks to wax synthesis. It has been found that gl₅ is a mutation which blocks elongation in the Step C₂₈–C₃₀. The mutation gl₄ exhibits no elongation block and could be blocked in the decarboxylation Step C₃₀–C₂₉. 0·1 mM TCA supplied in the culture solution of cauliflower seedlings affected the leaf surface by producing a glossy appearance similar to that induced by gl₃ and gl₄. At this concentration growth was not inhibited and the appearance of the plants was normal except for the surface wax. The amount of surface wax produced was about 40% of that in untreated seedlings on a leaf area basis. Slight, but significant changes in wax composition were noted, mainly involving a reduction in C₃₀ acids and aldehydes, a slight reduction (33–29%) in alkane content, and a marked difference in chain length composition of the alkanes with C₂₇ increased relative to C₂₉. Over a range of concentrations from 0·1–1 mM, TCA inhibited incorporation of label from acetate-1-¹⁴C into C₃₀ acids and aldehydes more than into C₂₈ at concentrations 0·4–0·8 mM while label tended to accumulate in C₂₄ and C₂₆ acids; thus elongation C₂₈–C₃₀ was especially sensitive to TCA. TCA also inhibited incorporation into primary alcohols and esters almost as much as into C₂₉ compounds. In spite of relatively specific effects on incorporation of label into longer chain lengths, the resulting block to C₃₀ synthesis is not sufficient to make much difference to the overall rate of C₂₉ synthesis. Both results of analysis of wax from whole plants and experiments with tissue slices in vitro indicated that the effect of TCA in reducing the glaucousness of the leaf surface is a combination of overall reduction of wax synthesis together with slight but significant changes in wax composition.     

Investigation of Brassica oleracea and Nasturtium officinale seeds for the presence of epithiospecifier protein

Cabbage seeds contain 5 glucosinolates and on autolysis produce, in particular, 1-cyano-2-3-epithiopropane. Watercress seeds contain 4 glucosinolates, but none capable of forming a cyanoepithioalkane. Cabbage exhibited behaviour commensurate with possession of an epithiospecifier protein (ESP) whilst watercress did not show any such activity.     

A chemotaxonomic study of Brassica oleracea with particular reference to its relationship to Brassica alboglabra

A chemotaxonomic study has been made of Brassica oleracea L., its horticultural varieties and Brassica alboglabra Bailey with reference to seed proteins, certain isoenzymes and isothcioyanates. The results of the investigation do not support separate species status in respect of B. alboglabra.     

Lipids of Pinus halepensis pollen

The total lipids of Pinus halepensis pollen were separated into individual classes of neutral and polar lipids and the components of each class were identified and determined quantitatively. Free fatty acids, waxes and triacylglycerols were found as the main constituents of neutral lipids and phosphatidylcholine and phosphatidylethanolamine of polar lipids. Glycerylether derivatives were detected in neutral and polar lipid fractions. Free and esterified volatile fatty acids were also found in pollen and its neutral lipid fraction.     

C-galactosylflavones from Polygonatum multiflorum

8-C-Galactosylapigenin and 6-C-galactosyl-8-C-arabinosylapigenin were isolated from the leaves of Polygonatum multiflorum (L.) All. Structural assignments for the latter compound were made on the basis of mass, CD and ¹³C-NMR spectra.     

Seasonal variation in the glucosinolate content of North American Brassica nigra and Dentaria species

Concentrations of glucosinolates in the leaves of the woodland cruciferous herbs Dentaria laciniata, D. diphylla and D. maxima declined during the 1974 growing season but generally equalled or exceeded the levels found in the foliage of Brassica nigra, a crucifer of neighbouring open habitats. The superior quality of Dentaria foliage for certain crucifer-feeding insects does not seem to result from an unusually low concentration of glucosinolates. The qualitative and quantitative compositions of glucosinolates in the leaves and rhizomes of D. maxima were found to be intermediate between the corresponding values for D. laciniata and D. diphylla. This finding is consistent with a hybrid origin for D. maxima.     

The effect of membrane stabilizers on phytochrome-controlled anthocyanin biosynthesis in Brassica oleracea

The promotion of anthocyanin synthesis in red-cabbage seedlings by 5 min exposure to R light is inhibited by subsequent application of CaCl₂. The stimulation of dark synthesis of anthocyanin by n-PrOH and by kinetin is also reduced by Ca²⁺ and by cholesterol, both of which are well known to stabilize cell membranes. By contrast, EDTA, which chelates Ca²⁺, promotes dark synthesis of anthocyanin. Assay of native Ca²⁺ extractable from seedlings immersed in EDTA demonstrates that R light exposure promotes a highly significant increase in extractable Ca²⁺. It is suggested that the molecular configuration of the phytochrome molecule affects the ability of a membrane to bind Ca²⁺ and that this in turn affects the permeability to substrates which are required for anthocyanin biosynthesis.     

Isolation and characterization of an iron-containing superoxide dismutase from a eucaryote, Brassica campestris

A cyanide-insensitive superoxide dismutase was purified from mustard leaves, Brassica campestris. The protein had a molecular weight of 41,000 and was composed of two equally sized subunits. Metal analysis revealed that the enzyme contained 1.6 g atoms of iron per dimer. The isolation of an iron-containing superoxide dismutase from mustard leaves represents the first report of this enzyme in a multicellular eucaryotic organism.     

Invertases of pollen of Haemanthus albiflos

Soluble and insoluble invertase occurs in dormant pollen of Haemanthus albiflos, with pH optima of 5·7 and 5·5 respectively. At their pH optima the activity of the soluble enzyme is 3·5-fold higher. After 2 hr germination the pH optimum of the insoluble invertase is increased to 6·0 and the activity is increased 2-fold while the activity of the soluble invertase is decreased by 26%.