Bile pigment synthesis in plants. Incorporation of haem into phycocyanobilin and phycobiliproteins in Cyanidium caldarium.

A procedure was developed whereby haem was taken up by dark-grown cells of the unicellular rhodophyte Cyanidium caldarium. These cells were subsequently incubated either in the dark with 5-aminolaevulinate, which results in excretion of phycocyanobilin into the suspending medium or incubated in the light, which results in synthesis and accumulation of phycocyanin and chlorophyll a within the cells. Phycocyanobilin was isolated from phycocyanin by cleavage from apoprotein in methanol. Phycocyanobilin prepared from phycocyanin or excreted from cells given 5-aminolaevulinate was methylated and purified by t.l.c. By using 14C labelling either in the haem or in 5-aminolaevulinate administered, haem incorporation into phycocyanobilin was demonstrated in both dark and light systems. Since chlorophyll a synthesized in the light in the presence of labelled haem contained no radioactivity, it was clear that haem was directly incorporated into phycocyanobilin and not first converted into protoporphyrin IX. These results clearly demonstrate phycocyanobilin synthesis via haem and not via magnesium protoporphyrin IX as has also been postulated.     

POLLEN MORPHOLOGY OF HAPLOPAPPUS AND RELATED GENERA (COMPOSITAE—ASTEREAE)

Pollen grains of Haplopappus and related genera in the subtribe Solidaginae from North and South America were examined by light and scanning electron microscopy. The grains are consistently tricolporate and echinate. Some genera can be distinguished by pollen size, spine length, and number of spine rows between colpi. Based on these characters, the divergence of Benitoa from other members of the subtribe, as indicated by its morphology and secondary chemistry, is supported. Additionally, the recently suggested absence of a close relationship between Pyrrocoma and Oonopsis is indicated by their contrasting pollen types. This study demonstrates the potential of pollen studies in distinguishing some taxonomic groups in the Astereae.     

Biosynthesis of the pyrimidinyl amino acid lathyrine by Lathyrus tingitanus L.

The biosynthesis of the pyrimidinyl amino acid lathyrine by seedlings of Lathyrus tingitanus L. was shown to be stimulated by uracil. [6(-14)C]Orotate, [2(-14)C]uracil and [3(-14)C]serine were incorporated into lathyrine; the incorporation of [6(-14)C]orotate was substantially decreased in the presence of uracil. Chemical degradation to locate the 14C incorporated from labelled precursors showed that 90% of the radioactivity incorporated into lathyrine from [3(-14)C]serine could be recovered in the alanine side chain. Over 80% of the radioactivity incorporated from [2(-14)C]uracil was shown to be located in C-2 of lathyrine. It is concluded that under the conditions studied, lathyrine arises from a preformed pyrimidine arising via the orotate pathway. Paradoxically, it was also possible to confirm previous reports that radioactivity from L-[guanidino-14C]homoarginine is incorporated into lathyrine and gamma-hydroxyhomoarginine. However, as homoarginine and gamma-hydroxyhomoarginine are also both labelled by [2(-14)C]uracil, it is suggested that they are products of the ring-opening of lathyrine and that reversibility of this process accounts, at least in part, for their observed experimental incorporation into lathyrine.     

Gene dose effect: intraband mapping of the LDH A locus using cells from four individuals with different interstitial deletions of 11p

The quantitative expression of LDH A was studied in hemolysates from four patients with different but overlapping interstitial deletions of the short arm of chromosome 11. Deficiency of LDH A was demonstrated in one patient, and the LDH A locus has been assigned to that segment of 11p for which this patient alone was deficient, i.e., to band 11p12 (region 11p1203 leads to 11p1208).     

The carotenoids of Flavobacterium strain R1560

The main carotenoid of Flavobacterium strain R1560 has been identified as (3R,3R)-zeaxanthin. Also present were small amounts of 15-cis-phytoene, phytofluene, -carotene (7,8,7,8-tetrahydro-, -carotene plus 7,8,11,12-tetrahydro-, -carotene), neurosporene, lycopene, -zeacarotene, -carotene, -carotene, -cryptoxanthin, rubixanthin, 3-hydroxy--zeacarotene and several apo-carotenals. Zeaxanthin production was inhibited by nicotine (10 mM), and lycopene and rubixanthin accumulated. The biosynthesis of zeaxanthin is discussed in terms of pathways and also of half-molecule reaction sequences. The presence of zeaxanthin may be a characteristic of a group of Flavobacterium species, and may thus be useful in the taxonomic classification of these organisms.     

An examination of the developing cotton fiber: Wall and plasmalemma

Summary The ultrastructure of developing cotton fibers has been examined using novel modifications of the techniques of surface replication, freeze-etching and thin-sectioning. The fiber surface was found to be coated with a lamellar cuticle, which is stretched and thinned as the fiber elongates. It is marked by bars which run parallel with the fiber long axis. Beneath the cuticle, the outermost microfibrils of the primary wall lie parallel with the fiber axis, while those adjacent to the plasma membrane are transverse. Primary wall microfibrils are present in bundles, disposed in left-handed and right-handed helices, which correspond with the fibrils observed optically. Microfibrils within bundles form in-phase waves, with wavelengths and amplitudes in the ranges 0.3–7 m and 0.01–0.1 m in primary and secondary walls respectively. As elongation proceeds bundles become displaced towards the cell axis. Microfibrils of the secondary wall, disposed around the cell as fast helices, are similarly bundled and wavy (though with a reduced amplitude). In surface-replicas, large (20–30 nm) granules are present on the cytoplasmic face of the wall which probably correspond with 20–40 nm low prominences visible on freeze-etch EF plasma membrane fracture faces. It is proposed that these may be microfibril-synthesizing centers. Plasma membranes fracture such that the membrane-associated-particles segregate 6040 between P and E fracture moieties, but the prominence and total number of these particles is reduced at the stage of secondary wall formation as compared with primary wall formation. Beneath the plasmalemma the axes of microtubules parallel secondary wall microfibril orientation. Cross-bridges, which stain heavily after glutaraldehyde/tannic acid fixation, link microtubules to plasma membrane. The use of butyl benzene to cement fragments of cotton fibers, employed in this work, may prove useful in other freeze-etch studies of long fibers which are readily ruptured during preparation.     

Properties and subcellular distribution of ribonuclease activity in Chlorella

RNase activity from Chlorella was partially purified. Two RNase activities were demonstrated, one soluble and the other ribosomal. The effects on ribonuclease activity of variations in pH and temperature, and of Mg²⁺, Na⁺, and mononucleotides were examined. The RNase activities (phosphodiesterases EC 3.1.4.23) were both endonucleolytic, releasing oligonucleotides, and cyclic nucleotide intermediates, but exhibited different specificities in releasing mononucleotides from RNA. The ribosomal activity released 3′-GMP, and after prolonged incubation 3′-UMP, but the soluble activity released 3′-GMP, 3′-AMP and 3′-UMP. Neither ofthe RNase preparations hydrolysed DNA, nor released 5′-nucleotides from RNA. Increased ribosomal RNase activity was related to dissociation of ribosomes, and latency of ribosomal RNase activity was demonstrated. The possible in vivo distribution of RNases is discussed.     

Copper as essential to wheat reproduction

Summary The role of Cu in promoting the reproductive phase of growth was examined using Thatcher spring wheat (Triticum aestivum L.). Plants were grown in limed Bladen soil (pH 5.3) in a controlled growth room. The plants were first harvested when five leaves had developed (day 23) and some top leaves of Cu-deficient (-Cu) plants had rolled (withertip) indicating a Cu deficiency. The leaves were shorter on Cu than Cu-sufficient (+Cu) plants, and fewer leaves developed. As they entered the reproductive growth phase, +Cu plants accumulated reducing sugars and reduced 2, 3, 5- triphenyltetrazolium chloride (TTC) in their stems, while Cu plants had lower concentration of the reducing sugars and very little TTC was reduced indicating a reduction in energy and in reducing capacity. Thin-layer-chromatography showed that top leaves of Cu plants contained higher concentrations of aspartic acid, alanine, and serine; and less aminobutyric acid than +Cu plants. Nitrate, P, and K concentrations were higher, and Ca and Cu were lower in Cu than in +Cu plants.Contribution of U.S. Department of Agriculture, Agricultural Research Service, Plant Stress Laboratory, Northeast Region, Beltsville, MD 20705 and North Central Region, Department of Agronomy, University of Nebraska, Lincoln, NE 68583 as Paper No.5187, Journal Series, Nebraska Agricultural Experiment Station.Contribution of U.S. Department of Agriculture, Agricultural Research Service, Plant Stress Laboratory, Northeast Region, Beltsville, MD 20705 and North Central Region, Department of Agronomy, University of Nebraska, Lincoln, NE 68583 as Paper No.5187, Journal Series, Nebraska Agricultural Experiment Station.     

Thermocouple psychrometer for in situ leaf water potential determinations

Summary An in situ thermocouple psychrometer was tested to determine its capability for tracking the changing leaf water potentials of aspen (Populus tremuloides Michx.) under controlled conditions and in the field. The results agreed with theoretical expectations — a sharp decline in leaf water potential during the photoperiods and recovery during the dark periods, each to progressively lower levels during a soil drying cycle. In the field, a mid-day pause in the declining leaf water potentials was noted and attributed to transient stomatal closure. Details of the construction and use of the assembly are given.The authors are plant physiologists, USDA Forest Service, Intermountain Forest and Range Experiment Station, Ogden, Utah 84401, stationed in Logan, Utah, at the Forestry Sciences Laboratory.