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Glandless cotton plants (Gossypium hirsutum L. cv. Coker 100) were subjected to the influence of high CO(2)-bicarbonate. The content of protein decreased with no accompanying increase in its degraded products. The decrease in protein was correlated with the low content of chlorophyll and also with the reduced activity of carbonic anhydrase. The initiation of these correlations coincided with the time when the control leaves contained the highest enzyme activity during leaf growth. The high concentration of bicarbonate directly restricted the rate of photophosphorylation and that of the Hill reaction in isolated chloroplasts. The amount of ATP in leaves treated in vivo also diminished. High CO(2) as bicarbonate, however, did not directly inhibit the activity of carbonic anhydrase in vitro.

Investigation was made for the effect of fluoride on plant growth, acid soluble nucleotides, and RNA in germinating corn seedling roots. Fluoride suppresses root growth as measured by changes in fresh weight. Column chromatographic analyses demonstrated that fluoride modifies ratios of acid soluble nucleotide species. The relative amount of nucleotides is altered mainly due to triphosphate nucleotides of which ATP is most accumulated. Paper chromatographic analyses showed that fluoride induces changes of RNA structure. The RNA is characterized by lowered relative content of cytosine and by increased ratio of cytosine to guanine. Adenine is depressed significantly only in the root tissue treated by the highest fluoride concentration.

Fluoride-polluted navel orange leaves, Citrus sinensis (Linn.) Osbeck, were fractionated into the subcellular components in hexane/carbon tetrachloride mixtures having various densities. Fluoride was determined at each fraction. Analyses were also made for the subcellular distribution of chlorophyll, nitrogen, and DNA to assess the extent of cross-contamination of each component. The fraction containing cell wall, nuclei, and partly broken cells apparently contained a major amount of fluoride. However, if allowance was made for the cross-contamination of chloroplasts and chloroplast fragments, the fraction of chloroplasts was found to be the site of the highest fluoride accumulation. When each particulate component was washed with water after drying, the combined washings contained more than 50% of the total fluoride of the isolated fractions. The usual method of subcellular fractionation with aqueous solvent shifted the major site of fluoride accumulation from the fraction of chloroplasts to that of the supernatant.

Cotton plants (cv. Coker 100) were exposed to a 14-hour dark period. Starch degradation occurred with no accumulation of sugars due mainly to translocation. Considerable amounts of starch degradation products however were detected from leaves after phloem transport was blocked. A minor component (10 to 25% of total starch) with a linear structure, amylose, was preferentially degraded, whereas the major multiple-branched component (about 80%), amylopectin, showed an increasing resistance to degradation with leaf age. This relationship was also shown by the decreasing iodine-binding capacity of unit starch with increasing leaf age. The structural resistance of amylopectin to enzymic dark degradation was one of the barriers to starch dissolution in cotton.

During cotton leaf development, starch accumulation was characterized by an initial rise to a maximum at the second to the fourth leaf from the apex. Then, starch content progressively decreased with leaf age. Starch accumulation was inversely related to the ratio of amylopectin to amylose. Differences between leaves in this ratio resulted from variations in both amylose and amylopectin levels. Fluctuations in amylose levels were more extreme than those of amylopectin. During the diurnal cycle, amylopectin was accumulated more than amylose in both young and old leaves during the day. During the night, amylopectin was degraded more than amylose in young leaves and vice versa in older leaves. The rate of change of the amylopectin to amylose ratio during the day was consistently higher than that during the night.

A procedure was developed for the separation and purification of amylose and amylopectin isolated from cotton leaves. Cotton leaves were homogenized in 0.02 molar phosphate buffer at pH 7.0 containing HgCl(2) plus toluene. Crude starch granules were collected by centrifugation and partially purified by treating with acetone and toluene. The starch granules were then dispersed in dimethylsulfoxide and precipitated with ethyl alcohol. The precipitate was suspended in boiling water. Amylose was separated from amylopectin and cell wall particles on a Sepharose 2B column and further purified with thymol and butanol. Amylopectin was then separated from the colloidal cell wall contaminants by its specific interaction with concanavalin A. Purities of starch components were verified by specific biochemical and enzymic tests in addition to their iodine-binding capacity. This procedure should also be suitable for purification of starch components from other plant sources.

Little attention has been given to the occurrence of exocellular hydrolytic enzymes associated with the roots of higher plants. Such enzymes, if present, could solubilize macromolecular constituents of organic soils, permitting uptake and utilization by the plant of the resultant low molecular weight organic compounds. The present work supports this concept by demonstrating that several exocellular hydrolytic enzymes are associated with the roots of intact corn seedlings. Distinction has been made between exocellular enzymes released into the culture media and enzymes apparently bound to the root surface. A previous paper from this laboratory (17) reviewed earlier studies and presented evidence for the occurrence of exocellular enzymes associated with the primary cell wall of corn coleoptiles. Of particular relevance to the present work is a report (36) of the secretion of peroxidase, acid phosphatase, indolylacetic acid oxidase, and amylase into the culture media bathing plant *tissue cultures. In addition, reports have appeared (2, 6, 14) of the occurrence of an exocellular invertase, possibly involved in sucrose metabolism by roots. In the present work it is reported that invertase, cellobiase, adenosine triphosphatase, pyrophosphatase, and nuclease activity is observed when appropriate substrates are added to nutrient solution bathing the roots of intact seedling corn plants. In addition, inveritase and nuclease are detected as soluble enzymes in the nutrient solution.