The accumulation of anthocyanin by clones and subclones from a cell suspension culture of wild carrot (Daucus carota L.) has been measured under standard conditions. Clones which accumulate low amounts of anthocyanin were shown, by recloning after maintenance by serial passage, to have become heterogenous and to contain cells with increased accumulation of anthocyanin. There appears to be a maximum amount of anthocyanin that clones can accumulate. Clones which accumulate the maximum amount of anthocyanin were shown by recloning after maintenance by serial passaging, to have become heterogenous and to contain many cells which accumulate less than the maximum possible amount of anthocyanin. When clones which accumulate the maximum amount of anthocyanin are maintained by serial passage, the decline in anthocyanin accumulation is different in different media. The results indicate that the changes in the ability of cells to accumulate anthocyanin involve no qualitative change in the genetic information of the cells, i.e., the changes are not the consequence of mutations.

Polymers of L-lactic acid are shown to promote plant growth . Dry weight of duckweed (Lemna minor L .) and corn (Zea mays L) was more than doubled when plants were grown in media containing the dimer of L-lactic acid, L-lactoyllactic acid . Higher polymers were equally effective at increasing plant biomass . Monomeric lactic acid and polymers of D-lactic acid showed no biological activity . Increased plant biomass was accompanied by increased chlorophyll accumulation and root growth . Promotion of chlorophyll accumulation and biomass may be due to increased ability to assimilate nutrients as plants treated with L-lactoyllactic acid showed no decrease in biomass when grown in medium that was growth limiting for control plants .Abbreviations . DP = degree of polymerization, with the degree indicated by the following subscript number ; LA = lactic acid .

With carrot cells grown in semicontinuous culture with phosphate as limiting nutrient. Dougall and Weyrauch (1980) found that the steady-state culture density was different at different dilution rates. They suggested that the yield constant for biomass was different at different dilution rates. Here the yield constant for biomass for PO(4) (3-), NH(4) (+), Mg(2+), and glucose-limited semicontinuous cultures has been measured directly at two dilution rates. The yield constants for PO(4) (3-), NH(4) (+), and Mg(2+) but not for glucose are different at the two dilution rates. The effects of pH and temperature on the biomass yield constant was measured to extend the number of system parameters examined. Biomass yield constant was changed little with change from 25 to 28 degrees C or from pH 4.2 to pH 5.5. The steady-state levels of anthocyanin were also measured. The responses of anthocyanin levels to the system parameters are different to the biomass responses. The data suggest that at different values of each of the system parameters, the composition and metabolic activities of the cells at steady state in semicontinuous cultures are different.

When carrot cell cultures, after growth in semicontinuous culture, were transferred to media containing excess nutrients, they grew at different rates. The growth rates were generally higher after semicontinuous culture at higher dilution rates. There appears to be a limit on dilution rate above which growth rate does not increase. These changes were also displayed by clones from the parental culture. The possibility that these changes in growth rate reflect a need for the cultures to adapt to their new conditions is discussed. The growth rates of the cultures is markedly increased at 25 degrees C compared with 22 degrees C with little further increase at 28 degrees C. Growth rate is altered by less than 20% when pH is changed from 4.5 to either 5.5 or 4.2. The rates of anthocyanin accumulation by the cultures were similar under all conditions tested except at 22 degrees C. They were larger than the rates of dry weight accumulation. In contrast, the amounts of anthocyanin accumulated in the clones and in the parental cultures grown at pH 5.5 instead of pH 4.5 were very different. The observations were interpreted as showing that the clones differ in the rate of metabolism but not in the rate of synthesis of anthocyanins and that at pH 5.5 the rate of metabolism of anthocyanins but not the rate of synthesis is higher than it is at pH 4.5. The use of semicontinuous cultures as a source of inoculum for batch cultures rather than as a source of biomass for extraction of chemicals is discussed.

Twenty-five different anthocyanin producing cultures of wild carrot were stored at-140" and recovered. In all CaSes , viable cultures were recovered which produced approximately the Same amount of anthocyanin as did similar cultures which had not been frozen and thawed. This is the first demonstration that plant cell cultures retain their ability to produce specific chemicals after cryogenic Storage. In addition, the data provided show that a reciprocal relationship between growth of cultures and their ability to produce specific chemicals does not always hold true.