Induction of anthocyanin accumulation in rose suspension-cultured cells by conditioned medium of strawberry suspension cultures

A conditioned medium (CM) prepared from suspended cultures of strawberry, Fragaria ananassa, which stimulated anthocyanin accumulation in cultured strawberry cells, was applied to the suspension-cultured cells of rose, Rosa hybrida sp which did not normally produce anthocyanin. When the rose cells were transferred into the CM, it induced anthocyanin formation and accumulation in the rose cells. It is suggested that the CM may be effective for inducing anthocyanin accumulation in cultured cells of other species. This revised version was published online in June 2006 with corrections to the Cover Date.     

Changes in PAL,CHS, DAHP synthase (DS-Co and Ds-Mn) activity during anthocyanin synthesis in suspension culture of Fragaria ananassa

The activity of 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase (DS-Mn, DS-Co), phenylalanine ammonia-lyase (PAL), and chalcone synthase (CHS) was monitored at various light intensities (dark, 8.88 μmol m⁻² s⁻¹, 88.8 μmol m⁻² s⁻¹) using a strawberry cell suspension culture. DS-Mn, PAL, and CHS were found to increase significantly (p>0.05) under light intensitie of 88.8 μmol m⁻² s⁻¹ compared to those of 8.88 μmol m⁻² s⁻¹ and dark. The activity of DS-Mn, PAL, and CHS were maximum at 88.8 μmol m⁻² s⁻¹. Anthocyanin content reached a maximum after 48–60 h of culturing at 88.8 μmol m⁻² s⁻¹. DS-Co showed greater activity than DS-Mn during cell culturing, but showed no correlation with anthocyanin production and light intensity. The CHS gene expression was continuous at a light intensity of 88.8 μmol m⁻² s⁻¹. This revised version was published online in June 2006 with corrections to the Cover Date.     

Impaired Ca2+ store functions in skeletal and cardiac muscle cells from sarcalumenin-deficient mice

Sarcalumenin (SAR), specifically expressed in striated muscle cells, is a Ca2+-binding protein localized in the sarcoplasmic reticulum (SR) of the intracellular Ca2+ store. By generating SAR-deficient mice, we herein examined its physiological role. The mutant mice were apparently normal in growth, health, and reproduction, indicating that SAR is not essential for fundamental muscle functions. SAR-deficient skeletal muscle carrying irregular SR ultrastructures retained normal force generation but showed slow relaxation phases after contractions. A weakened Ca2+ uptake activity was detected in the SR prepared from mutant muscle, indicating that SAR contributes to Ca2+ buffering in the SR lumen and also to the maintenance of Ca2+ pump proteins. Cardiac myocytes from SAR-deficient mice showed slow contraction and relaxation accompanied by impaired Ca2+ transients, and the mutant mice exhibited a number of impairments in cardiac performance as determined in electrocardiography, ventricular catheterization, and echocardiography. The results obtained demonstrate that SAR plays important roles in improving the Ca2+ handling functions of the SR in striated muscle.     

Conditioned medium from heterogeneous plants (rose and grape) on cell growth and anthocyanin synthesis of Fragaria ananassa

Rose and grape cells were cultured under 800 lux for 1 week, and these broths were used as conditioned medium for stimulate anthocyanin synthesis of strawberry suspension cells. Anthocyanin synthesis in strawberry cells was enhanced to produce 1000 ug/g cells after 2 days of culturing in the conditioned medium prepared from white grape callus which does not produce anthocyanin, and the enhancement was significantly (p < 0.05) greatest and it was 5 fold greater than that of the control medium. Stimulation by conditioned media also showed a concentration-dependent response for anthocyanin synthesis, although there was only a slight difference (4.0-4.8 g) in cell growth among the heterogeneous conditioned media after 2 days of culturing. However after 5 days, cell growth using 50% of conditioned media from red grape callus was 14.6 g cell weight and 15 g in the control.     

Changes of anthocyanin composition by conditioned medium and cell inoculum size using strawberry suspension culture

Summary Production of anthocyanin was greater in conditioned medium than in LS medium. The highest production was obtained after 2 days culturing, and the value reached 1100 g/g cell, which was about twice that of LS medium cultured in each cell inoculum. The highest anthocyanin content was obtained with a 1g cell (fresh weight) inoculation using conditioned medium. The percentage of the main anthocyanin, peonidin-3-glucoside, increased in accordance with the decrease in cell inoculum size, contrary to the results obtained for cyanidin-3-glucoside. This tendency was more clearly observed in LS medium than in conditioned medium.     

Preparation of conditioned medium to stimulate anthocyanin production using suspension cultures of Fragaria ananassa cells

A conditioned medium (CM) prepared from strawberry suspension cultures greatly stimulated anthocyanin accumulation. CM separated by dialysis membrane showed a significant increase (p 0.05) in anthocyanin synthesis at a fraction smaller than 10,000 Da. The stimulation by CM was eliminated when the CM was treated with alkali.