Increased protoplast yield from oat leaves and bean internodes by non-injurious mechanical perturbation

Summary A simple new technique has been developed to greatly increase the yield of protoplasts from plant organs without injury to the plant. Mechanical perturbation (MP) by non-stressful rubbing of oat leaf segments and bean internodes yielded ten to twenty times more viable protoplasts than did controls. The increase in protoplast yield due to MP is best manifested, if the organs are excised and transferred to the cellulytic enzymes immediately after MP is given to the intact organ. The enzymes begin digesting from the lower end of the bean internodes and proceed acropetally. Vacuum infiltration of control oat leaf segments for 15 min with enzyme solution resulted in increased yield but less than due to MP. Increased levels of calcium (10 mM) in the medium decreased the yield of protoplasts from both control and MP-treated plant organs. EGTA significantly increased the yield of protoplasts from control oat leaf segments and marginally over that found in the control bean internodes. Cycloheximide increased the yield of protoplasts from oat leaf segments but not from bean internodes. It is suggested that MP may increase the susceptibility of cell wall polymers to cellulytic enzymes by reducing calcium cross linking. MP is thus a tool for increasing the yield of protoplasts from plant organs without causing injury.Abbreviations CHI cycloheximide - EGTA ethyleneglycol-bis-(ß-aminoethyl ether)-N,N-tetraacetic acid - FDA fluorescein diacetate - MP mechanical perturbation     

Studies on light absorption and photochemical activity changes in chloroplast suspensions and leaves due to light scattering and light filtration across chloroplast and vegetation layers

An experimental analysis is presented concerning the effect on relative light absorption by the two photosystems caused by (a) a highly light scattering environment (the detour effect) and (b) light filtration across successive chloroplast layers (the light attenuation effect). Both suspensions of isolated chloroplasts and leaves were employed.It is concluded that within a single spinach leaf these phenomena are likely to lead to only rather small increases in relative photosystem I absorption and activity with respect to photosystem II and will thus not exert a significant effect on non cyclic electron transport. On the contrary when light is filtrated across successive vegetation layers (shade light) significant increases in the relative PSI absorption and activity may be encountered.It is determined that the detour effect in mature leaves from a variety of plants increases overall photosynthetically useful light absorption by 35–40%.Abbreviations F_M maximal fluorescence - LHCP₂ light-harvesting chlorophyl a/b protein complex II - Q_A-primary quinone acceptor of photosystem II     

Flavonoid aglycones from the leaf surfaces of someLabiatae species

The distribution of excreted flavonoid aglycones within the familyLabiatae was studied and differences were found, especially in the A-ring substitution patterns. Thus, 5,7-dihydroxy-6-methoxyflavones with substituted B-rings are characteristic of species ofSalvia (sect.Salvia),Rosmarinus andOcimum; 5,7-dihydroxy-6,8-dimethoxyflavones occur only inOcimum and 5,6-dihydroxy-7,8-dimethoxyflavones inThymus and related species. Members of the two subfamiliesLamioideae andNepetoideae produce exudate flavonoids, but some genera are devoid of these compounds. There is a correlation between the habitat where the plant grows and production of these compounds, the species from (semi-)arid habitats being those which generally accumulate external flavonoids.     

Delimitation ofPelargonium sect.Glaucophyllum (Geraniaceae)

Pelargonium otaviense Knuth andP. spinosum Willd. are excluded from sect.Glaucophyllum, whileP. grandiflorum (Andr.)Willd.,P. patulum Jacq. andP. tabulare (Burm. f.)L'Hérit. of sect.Eumorpha are included. Sect.Glaucophyllum is characterized by green to glaucous vegetative organs and zygomorphic white to pink corolla with five narrow petals. All the species have an identical pollen and chromosome morphology, the same basic chromosome number (x = 11) and similar flavonoid patterns. A close relationship between sect.Glaucophyllum and sect.Pelargonium is indicated by the occurrence of natural hybrids and concordant characters. Isorhamnetin and luteolin have been detected in the genus for the first time.     

广西百色盆地更新世樟科两种植物角质层研究

本文描述的具角质层的两块叶化石,产自广西百色盆地更新统长蛇岭组。经与现代植物的角质层对比研究后,确定这两块叶化石分属于樟科的两属两种,即油丹(近似种)(Alseoda-phne cf.hainanensis Merr.)和紫楠(近似种)[Phoebe cf.sheareri(Hemsl.)Gamble]。研究结果表明,角质层在鉴定被子植物化石中具有可靠的价值。     

Effect of chloramphenicol and lincomycin on chloroplast DNA amplification in greening pea leaves

The light-induced increase in chloroplast DNA was not inhibited by two inhibitors of protein synthesis on 70S polysomes, chloramphenicol and lincomycin, in greening pea leaves. The changes in chloroplast DNA were observed by fluorescence microscopy and measured by hybridization to specific cloned probes. The results suggest that the light-induced increase in chloroplast DNA proceeds without de novo protein synthesis in the chloroplast, in agreement with those with mutants and cultured leaf tissue.     

Identification of a calmodulin-stimulated (Ca2++ Mg2+)-ATPase in a plasma membrane fraction isolated from maize (Zea mays) leaves

Plasma membranes were isolated from light-grown, 14-day-old maize leaves ( Zea mays L . cv. Golden Cross Bantam) using aqueous two-phase partitioning. The plasma membrane (PM) fraction contained < 0.3% of the total chlorophyll, < 0.2% of the mitochondrial marker enzyme activity, minimal contamination by endomembranes and 34% of the total PM.
A calmodulin-stimulated (Ca2++ Mg2+)-ATPase was identified in the PM-enriched fraction. The Ca2++ calmodulin stimulation was dependent on Mg2+, saturated at ca 25 μM total Ca2+, had a pH maximum at 7.2 and was maximally stimulated by 600 n M bovine brain calmodulin. The stimulation was not greatly affected by the anion present and showed a divalent cation specificity of Ca2+ > Sr+2 ± Mn+2 > Co2+± Cu2+ > Ba2+. The napthalenesulfonamide W7, an antagonist of calmodulin action, completely inhibited the calmodulin stimulation at 175 μM , while the less active analogue W5 was ineffective at this concentration. La3+, an inhibitor of PM Ca2+ transport, showed a 50% inhibition of calmodulin-stimulated ATPase activity at ca 200 μM . Taken as a whole, these data demonstrate the presence of a calmodulinstimulated, (Ca2++ Mg2+)-ATPase on the cytoplasmic surface of the plasma membrane of maize leaf cells.     

Phytochemical and morphological support for the existence of two species inMonoclea (Hepaticae)

Recognition of two different species in the liverwort genusMonoclea Hook. (monotypic orderMonocleales), viz.M. forsteri Hook. in New Zealand andM. gottschei Lindb. in the New World, is supported by characteristics of the sporophyte, antheridial receptacle and secondary metabolites.M. gottschei produces the greatest variety of flavonoids and the largest amount of bisbibenzyls ever encountered in a liverwort. In contrast,M. forsteri is poor in secondary metabolites. Two allopatric subspecies are recognized inM. gottschei, based on characteristics of the antheridial receptacle: subsp.gottschei in Chile (Valdivian region, Juan Fernandez Is.) and subsp.elongata Gradst. & Mues, subsp. nova, in tropical America. The exclusive occurrence inMonoclea of glucuronide and galacturonide flavone glycosides and the fact that capsule dehiscence may take place before full elongation of the seta are new arguments in support of the placement ofMonocleales in theMarchantiidae. Publication Nr. 43 of the Arbeitskreis Chemie und Biologie der Moose, Universität des Saarlandes, Saarbrücken. This paper is dedicated to DrElla O. Campbell, Massey University, Department of Botany and Zoology, New Zealand on the occasion of her 80th birthday.     

Salicyloylaspartate as an endogenous component in the leaves of Phaseolus vulgaris

An amide conjugate of o-methoxybenzoic acid and aspartic acid has been isolated from bean leaves. After extraction and methylation of plant material, this compound was isolated as two isomeric monoethyl monomethyl esters. The ethylation of the aspartyl carboxyl groups was shown to be a likely result of an extraction procedure utilising acidified ethanol, the methylation of the aromatic hydroxy of the methoxy group to be due to the derivatisation procedure. Studies with pentafluorobenzylation confirmed that the endogenous compound is o-hydroxybenzoylaspartate.     

Absolute configurations of pittosporatobiraside A and B from Pittosporum tobira

The absolute configuration at C-12 of pittosporatobiraside A and B isolated from the leaves of Pittosporum tobira was determined to be S on the basis of the exciton chirality of their dibenzoate derivative. The structures of the two glycosides were thus established to be (1S,9S,10S,11S,12S,14R,16R)-12-[(Z)-2-methyl-1-oxo-2-butenyl]-6,14-dimethyl-2-methylene-9-(1-methylethyl)-15,17-dioxatricyclo[8.7.0.0^11,16]heptadec-5-en-13-one and (1S,9S,10S,11S,12S,14R,16R)-12-(3-methyl-1-oxo-2-butenyl)-6,14-dimethyl-2-methylene-9-(1-methylethyl)-15,17-dioxatricyclo [8.7.0.0^11,16]heptadec-5-en-13-one, respectively.