LIGHT-INDUCED FORMATION OF ASCORBIC ACID IN ISOLATED CHLOROPLASTS

Investigations were made to determine the nature of a reducingsubstance which was formed on illuminating a reaction systemconsisting of chloroplast and cytoplasmic fraction of spinachleaves. From the results of spectrophotometric examinationsit was concluded that the photoproduct in question, or at leastits major portion, was ascorbic acid. The precursor of ascorbicacid, which was found to be heat unstable, was contained inthe cytoplasmic fraction. (Received October 12, 1960; )     

Fractal Analysis of Plant Root Systems

The morphology of root systems of crop plants was analysed byfractal geometry using an image processing system. Results indicatethat these root systems have a fractal structure (D; 1.48     

Structure and Role of the Five Glycopeptides of Human IgM Immunoglobulins

Carbohydrate is attached at five sites in the constant sequence region of the µ heavy chain of human IgM. The oligosaccharides are of two kinds, simple and complex and affect the conformation and properties of macroglobulins.     

Cell Fusion and Its Application to Genetic Analysis of Development in Dictyostelium discoideum

Dictyostelium cells were fused by a modification of the polyethylene glycol method of Kuhn and Parish. In the modified method Tricine buffer and Concanavalin A were used in place of Ca⁺⁺. The efficiency of genetic complementation through cell fusion was about 10 times higer by the modified method than by the original method with glycine buffer and Ca⁺⁺. Complementation between developmental mutants without any selectable growth character was clearly detected by the modified system, at efficiencies of about 1 in 10–20 surviving cells.     

A simple method to score single nucleotide polymorphisms based on allele‐specific PCR and primer‐induced fragment‐length variation

We describe a simple protocol to genotype single nucleotide polymorphisms (SNPs), which combines allele‐specific polymerase chain reaction (PCR) with fragment‐length analysis. Three primers are used in the PCR: two allele‐specific forward primers with a length‐difference and one reverse primer. The forward primers induce a length‐difference between the SNP‐variants, which can be assessed with standard fragment‐length analyses. We designed primers for 21 SNPs, and codominance was achieved for 76% of these SNPs. An inexpensive and flexible laser‐detection scoring protocol can be achieved with multiplex scoring and by incorporating the M13(‐21) genotyping method.     

A stress‐inducible sulphotransferase sulphonates salicylic acid and confers pathogen resistance in Arabidopsis

Sulphonation of small molecules by cytosolic sulphotransferases in mammals is an important process in which endogenous molecules are modified for inactivation/activation of their biological effects. Plants possess large numbers of sulphotransferase genes, but their biological functions are largely unknown. Here, we present a functional analysis of the Arabidopsis sulphotransferase AtSOT12 (At2g03760). AtSOT12 gene expression is strongly induced by salt, and osmotic stress and hormone treatments. The T‐DNA knock‐out mutant sot12 exhibited hypersensitivity to NaCl and ABA in seed germination, and to salicylic acid (SA) in seedling growth. In vitro enzyme activity assay revealed that AtSOT12 sulphonates SA, and endogenous SA levels suggested that sulphonation of SA positively regulates SA production. Upon challenging with the pathogen Pseudomonas syringae, sot12 mutant and AtSOT12 over‐expressing lines accumulate less and more SA, respectively, when compared with wild type. Consistent with the changes in SA levels, the sot12 mutant was more susceptible, while AtSOT12 over‐expressing plants are more resistant to pathogen infection. Moreover, pathogen‐induced PR gene expression in systemic leaves was significantly enhanced in AtSOT12 over‐expressing plants. The role of sulphonation of SA in SA production, mobile signalling and acquired systemic resistance is discussed.     

Carapace formation of the podocopid ostracode Semicytherura species (Crustacea: Ostracoda)

Details of ostracode carapace structures were examined by SEM and TEM. The podocopine ostracode Semicytherura kazahana has major ridges on the carapace surface and develops its prismatic layer inside the adult carapace. Electron microscopy at the final molt reveals that the major ridges arise from the highly dense formation of pits within the underlying swollen epidermis, and that disappearance of the epidermis in the presumptive area of the prismatic layer occurs after the calcification of the outer lamella cuticle, and just before synthesis of the membranous layer. These facts suggest that the formation of the carapace in Semicytherura takes place via a more complex process than that of the other podocopid ostracodes.     

Temporal photosynthetic carbon isotope signatures revealed in a tree ring through 13CO2 pulse-labelling

Using a combined method of pulse-labelling trees and analysing detailed distribution of ¹³C tracer within tree rings, we studied how photo-assimilates incorporated on a given day are then distributed in a tree ring. A branch of a 4-year-old Cryptomeria japonica D.Don tree growing in Tsukuba, Japan was pulse-labelled with non-radioactive ¹³CO₂ on two occasions: 29 May 2001 and 18 September 2001. Two discs were cut from the stem on 4 March 2002, one immediately under and the other 0.5 m below the branch and put through high-resolution δ ¹³C analysis. δ ¹³C peaks were observed in both the earlywood and latewood of the concerned tree ring, corresponding to each pulse-labelling date. The earlywood peaks was broader than the latewood peaks, possibly reflecting seasonal variation of the width of wood developing zone. Half-widths of the peaks were measured and used as indicators for the potential time resolution of tree-ring isotope analysis. The half-widths of the peaks indicated a time resolution no finer than 8.7–28 and 33–42 d in the early and latewood, respectively. Holocellulose extraction yielded only a slight change to the shape of the δ ¹³C peaks. ¹³C tracer pulse-labelled in May and September reached tangentially different locations in the lower disc, suggesting a seasonal change in the pathway of carbohydrates. Local consumption of spring assimilates and long-distance downward transport of autumn assimilates were also suggested.     

Arabidopsis thaliana ASN2 encoding asparagine synthetase is involved in the control of nitrogen assimilation and export during vegetative growth

We investigated the function of ASN2, one of the three genes encoding asparagine synthetase (EC 6.3.5.4), which is the most highly expressed in vegetative leaves of Arabidopsis thaliana. Expression of ASN2 and parallel higher asparagine content in darkness suggest that leaf metabolism involves ASN2 for asparagine synthesis. In asn2‐1 knockout and asn2‐2 knockdown lines, ASN2 disruption caused a defective growth phenotype and ammonium accumulation. The asn2 mutant leaves displayed a depleted asparagine and an accumulation of alanine, GABA, pyruvate and fumarate, indicating an alanine formation from pyruvate through the GABA shunt to consume excess ammonium in the absence of asparagine synthesis. By contrast, asparagine did not contribute to photorespiratory nitrogen recycle as photosynthetic net CO₂ assimilation was not significantly different between lines under both 21 and 2% O₂. ASN2 was found in phloem companion cells by in situ hybridization and immunolocalization. Moreover, lack of asparagine in asn2 phloem sap and lowered ¹⁵N flux to sinks, accompanied by the delayed yellowing (senescence) of asn2 leaves, in the absence of asparagine support a specific role of asparagine in phloem loading and nitrogen reallocation. We conclude that ASN2 is essential for nitrogen assimilation, distribution and remobilization (via the phloem) within the plant.