Osmium tetroxide footprinting of a scaffold attachment region in the maizeAdh1 promoter

Osmium tetroxide (OsO₄) reacts with the thymine residues of double-stranded DNA, but thymines that are unpaired or under torsional stress are hyperreactive. Although OsO₄ hyperreactivity has been primarily utilized to identify Z-DNA structures in supercoiled plasmids, OsO₄ will also identify other torsional perturbations of DNA. In this study, OsO₄ was used to footprint an AT-rich region (between –780 and –500) of the maizeAdh1 promoter. Hyperreactive sites were identified bothin vitro andin vivo in an area that coincides with AT motifs similar to those found in scaffold attachment regions. Further, the region of OsO₄ hyperreactivity lies within a fragment of DNA that is associated with the nuclear scaffold in histone-depleted nuclei.     

Cloning,sequence analysis and expression of a cDNA encoding active phosphoenolpyruvate carboxylase of the C3 plant Solanum tuberosum

A cDNA coding for phosphoenolpyruvate carboxylase (PEPC) was isolated from a cDNA library from Solanum tuberosum and the sequence of the cDNA was determined. It was inserted into a bacterial expression vector and a PEPC^- Escherichia coli mutant could be complemented by the cDNA construct. A functional fusion protein could be synthesized in E. coli. The properties of this PEPC protein clearly resembled those of typical C3 plant enzymes.     

Arrest of somatic embryo development in grapevine: histological characterization and the effect of ABA,BAP and zeatin in stimulating plantlet development

In an effort to understand the causes of arrest of somatic embryo development, generally observed in grapevine (Vitis sp.), histological studies were undertaken, using two cultivars (CH76 and 41B) which differ in their ability to develop into plants. Embryos with a high conversion rate (70%; CH76) formed a well-structured and functional shoot apex between two thread-like cotyledons. In contrast, embryos with a low conversion rate (10%; 41B) formed a normal root apex but lacked a well-structured shoot apex and developed a wide range of aberrant forms in the intercotyledonary area: uncontrolled cellular proliferation, formation of adventitious buds, over-growth of cotyledonary or leaf meristems. ABA increased the conversion rate of 41B embryos from 10% to 20%, but failed to improve embryo morphology. Zeatin and BAP promoted growth of 41B somatic embryos, but generated a high level of abnormalities and failed to improve conversion rate. Applied in combination with ABA, these PGRs increased the frequency of cotyledonary embryos, but decreased the conversion rate.     

Fertile plants regenerated from suspension culture-derived protoplasts of an indica type rice (Oryza sativa L.)

Calluses were induced from immature embryos of an indica type rice and finely dispersed cell suspension cultures were initiated from the callus using modified AA medium (S₁ medium). The suspension cultures were maintained alternatively (1–2 passages in each medium) in S₁ medium and S₂ medium, the latter containing KNO₃, NH₄NO₃, proline and glutamine as nitrogen source. Protoplasts of high quality were isolated form suspension cells cultured in S₂ medium supplemented with ABA. Embedding the protoplasts in agarose blocks containing NH₄NO₃-free modified KM8P(PM₁) medium and immersing the blocks in NH₄NO₃-containing modified KM8P(PM₃) medium were most effective for obtaining protoplast division and callus formation. The protoplast-derived calluses were precultured in potato extract-aand/or ABA-containing N₆(D₁, D₂ or D₃) media and many embryo-like structures were formed. These structures developed into plantlets after being transferred to N₆ differentiation (D₄) medium. The regenerated plantlets grew into mature plants and beard seeds normally.Abbreviations AA medium amino acids based medium - ABA abscisic acid - BA benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - DF division frequency - IAA indoleacetic acid - KIN kinetin - NAA naphthaleneacetic acid - PE planting efficiency     

The Casparian Strip of Clivia miniata Reg. Roots: Isolation,Fine Structure and Chemical Nature*

The root endodermis of Clivia miniata Reg. was successfully isolated using the cell wall degrading enzymes cellulase and pectinase. The enzymes did not depolymerize those regions of the primary cell walls of anticlinal endodermal root cells where the Casparian strips were located. Since the endodermis of C. miniata roots remained in its primary developmental state over the whole root length, endodermal isolates essentially represented Casparian strips. Thus, sufficient amounts of isolated Casparian strips could be obtained to allow further detailed investigations of the isolates by microscopic, histochemical and analytical methods. Scanning electron microscopy revealed the reticular structure of the Casparian strips completely surrounding the central cylinder of the roots. Whereas in younger parts of the root only the anticlinal cell walls of the endodermis remained intact in the isolates, in older parts of the root the periclinal walls also restricted enzymatic degradation due to the deposition of lignin. Extracts of the isolates with organic solvents did not reveal any wax-like substances which might have been deposited within the cell wall forming a transport barrier, as is the case with cutin and suberin. However, several histochemical and analytical methods (elemental analysis and FTIR spectroscopy) showed that the chemical nature of the Casparian strips of C. miniata roots can definitely be a lignified cell wall. These findings are in complete agreement with studies carried out at the beginning of this century on the chemical nature of the Casparian strips of several other plant species. The implications of these results concerning apoplasmatic transport of solutes and water across Casparian strips are discussed.     

Size-dependent reproduction in Australian alpine Ranunculus

Abstract The effect of plant size on reproduction in four species of alpine Ranunculus (R. muelleri, R. dissectifolius, R. graniticola and R. niphophilus) was investigated in two sites over two seasons in the field on a total of 190 plants. The effects of plant size (number of leaves) and number of flowers on the number of anthers, ovules and seed per flower and per plant were determined. There was a positive relationship between several measures of reproduction and plant size in all four species, indicating that reproduction is size-dependent. All the results indicate that the main factor controlling the amount of seed produced by alpine Ranunculus is the size of the plant. Specifically, bigger plants produced more seed by producing more flowers, not by producing more ovules per flower, or higher seed set per flower. Correspondingly, bigger plants produced more anthers by producing more flowers, rather than by producing flowers with more anthers. The total number of seeds produced by a plant was directly proportional to plant size in the four species. Therefore, reproductive effort should not vary with plant size in the four species.     

Flower nectar of an autogamous perennialRorippa indica as an indirect defense mechanism against herbivorous insects

This report shows that one of the most important roles of the flower nectar of an autogamous perennialRorippa indica (L.) Hieron is as an attractant for employing some ant species as a defense against herbivorous insects. The plant has flowers from spring to early winter. Its flower nectar is frequently stolen by some ant species (hereafter cited as ants) which also feed on small herbivorous insects on the plant. Internations among the tritrophic levels (R. indica, herbivores, ants) were experimentally examined and the followings became clear. (1) Ants were attracted toR. indica in search of its flower nectar. (2) The gradual secretion of flower nectar seemed to detain ants on the plant. (3)Pieris butterfly lavae were the major herbivores onR. indica and were potentially harmful to the plant. (4) The presence of ants reduced the survival rate ofP. rapae larvae onR. indica. (5) The presence of ants reduced the feeding damage toR. indica. (6) The disadvantage of nectar use by ants seemed to be minimal for the plant since the ants did not disturb the other flower visitors. These facts suggest a mutualistic relationship betweenR. indica and ants. That is, the flower nectar serves as an indirect defense against herbivorous insects.     

Extra-weak chemiluminescence of drugs XV. Method for determining stability of Toki-shakuyaku-san extract granules

The purpose of the study was to evaluate the quality of Toki-shakuyaku-san extract granules (TJ-23) using chemiluminescence (CL). A linear relationship was obtained between the log value of the CL of TJ-23 and the reaction temperature. An excellent correlation (r= 0.999) was found between the slope of this curve (ΔA) and the colour intensity due to the browning reaction occurring at the early stage of the Maillard reaction.