Raffmose saccharides occur as storage carbohydrate in seeds of legumes (14,15) and other species (4,15,24) and in vegetative tissues of a wide range of plants (6,22, 30). In the Cucurbitaceae, stachyose appears to be the predominant transport sugar (12, 28, 29) rather than sucrose (31); however, raffmose and small amounts of verbascose also may be translocated (12). There is evidence that sucrose is also a transport sugar in some of these species (12, 28 (9,25). In the peduncle, stachyose is metabolized to sucrose which apparently enters the fruit and is further metabolized to glucose and fmctose (9, 20).The recent report of raffinose saccharides in the mature seeds of C. sativus 'Calypso' (11) suggests that in situ synthesis of these sugars occurs in the developing seed, particularly since these sugars were not detected by paper chromatography in the fleshy portions of the cucumber fruit (20). There is the possibility, however, that this method of analysis failed to detect these sugars if they were restricted to the vascular tissues of the fruit. They might have been diluted by the high levels of glucose and fructose in the nonvascular tissues.This study was conducted to determine if in situ synthesis is responsible for the appearance of raffinose and stachyose in maturing cucumber seeds. To answer this question, changes in carbohydrate levels within the mesocarp, endocarp, and seed of maturing cucumber fruit were measured. In addition, galactinol synthase activity, the first unique enzyme in raflmose saccharide biosynthesis (7,10,11,16,19), and the sugars in funiculi were determined. In this study, decrease in endocarp pH was used as an index of fruit maturation (21). To determine if this was an adequate estimate of maturity, a separate study was conducted comparing this decrease in pH with a loss in peel Chl. Also, the organic acids malate and citrate were assayed from endocarp tissue during fruit maturation.
MATERIALS AND METHODSPlant Material and Sampling Procedures. Cucumber (Cucumis sativus L.) fruit utilized in these studies were harvested from plants of cv Calypso, grown in field plots in Raleigh (except as noted below). For sugar and starch analysis offruit tissues, three separate harvests were made during the growing season. Fruit were hand harvested in early morning, washed, and separated into nine different categories based on the pH of the endocarp tissue (21). These pH categories ranged from 5.75 to 3.50 in 0.25 pH unit increments. Each fruit was cut crosswise at a point approximately one-third of the length of the fruit proximal to the blossom end. A pH electrode was inserted into the center of this one-third section, the pH recorded, and this section was discarded. The twothirds section was immediately chilled in ice until processed. Three fruit were collected for each pH category. All further operations were performed at 0°C.Endocarp tissue and seeds were scooped from the three fruit in each pH category. Endocarp tissue refers to the gelatinous tissue surrounding the seeds and includes the ...