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Energy dispersive X-ray microanalysis has been used to study the binding of two inhibitors of hatching in Globodera rostochiensis, ruthenium red and lanthanum, to a calcium binding site on the eggshell. Eggs were treated with either buffered ruthenium red or buffer only, and the levels of ruthenium and calcium that could be detected from the surface of eggshells were measured before and after repeated etching in a plasma oven. Counts for ruthenium increased during etching to a maximum after 8–12 min, just before the eggshell was eroded from the juvenile. The calcium levels on eggshells binding ruthenium red were suppressed relative to those from buffer. Similar experiments with lanthanum showed a significant increase in this inhibitor after 10 min etching and an accompanying suppression of calcium levels. The nature of the binding site was probed by measuring ruthenium red binding after pretreatment with selected enzymes. A non-specific protease, pronase, reduced the subsequent binding of this inhibitor but the site was resistant to trypsin in the presence of calcium. Neuraminidase also had an effect suggesting that the protein is a sialoglycoprotein. Phospholipase A2 but not Phospholipase C influenced the subsequent binding of ruthenium red. Phospholipase A2 caused a loss of hatching ability and the degradation of a phospholipid from the eggshell. Ruthenium red binding was also reduced by treatment of eggshells with detergents or sonication. Apparently a calcium-binding sialoglycoprotein forms part of the integral proteins of an eggshell membrane located on the inner surface of the eggshell. 相似文献
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3-(3,5-Dichlorophenoxy)butyric acid and 2-(3,5-dichlorophenoxy)ethylamine were both shown to exert pronounced dwarfing effects on tomato seedlings. Examination of extracts of the treated plants revealed the conversion of these compounds to 3,5-dichlorophenoxyacetic acid (3,5-D) and it is concluded that the physiological activity shown in each case depends on such a conversion to 3,5-D within the tissues. 相似文献