Ultrastructural investigation has shown the presence of bacteria in the perinuclear cisternae of Chrysosphaerella brevispina. Transmission of the bacteria was shown to take place during cell division. The presence of intracellular bacteria have been reported in a wide variety of algal classes. The Chlorophyceae until recently have had more reports than any other group of algae (Mishra, 1969; Kochert and Olson, 1970; Burr and West, 1970; Turner and Freidman, 1974; Lee and Kochert, 1976; Dawes and Lohr, 1978; Colombo, 1978). Observations of bacteria in other algal classes involve the Dinophyceae (Silva, 1959, 1962, 1963, 1967, 1978; Gold and Pollinger, 1971; Dodge, 1973; Lucas, 1982), Chloromonadophyceae (Heywood, 1978), and recently the first reports in the Cyanophyceae (Wujek, 1979), Xanthophyceae (Ott, 1979) and the Prasinophyceae (Wujek et al., 1982). The Chrysophyceae rank third in reports of endosymbiotic bacteria documented by electron microscopy (Belcher, 1969; Swale, 1969; Hibberd, 1971; Wujek, 1978; Wujek and Kristiansen, 1978). This investigation reports bacteria in the perinuclear space of Chrysosphaerella brevispina (Chrysophyceae). The only previous record of bacteria associated with the nucleus is Leedale's (1969) observation of endonuclear bacteria in the Euglenophyceae. MATERIALS AND METHODS Chrysosphaerella brevispina Korshikov was collected from ponds in the Lake Itasca, Minnesota region. The species was fixed, prepared, and examined by electron microscopy as previously described (Wujek, 1969). Sections were examined with the Philips EM 300 electron microscope. OBSERVATIONS AND DISCUSSION A number of the cells possessed bacteria in the expansion of the perinuclear cisternae (Figs. 1-3). No swelling of the perinuclear space was observed in This content downloaded from 157.55.39.212 on Thu, 09 Jun 2016 06:24:57 UTC All use subject to http://about.jstor.org/terms VOLUME 87, NUMBERS 1-2 33 o . ;. . . ,, . . .;ii .. , . t,, ', ? ? .. ..,, . -.-. , ril ' J.. ?? r?i ,:I ::i,._', O il, . ,.; , :;.,, o i '? i-. . . .. . ? , !,.,,, 7.l ? , -. . -. , .; ! .. . . : , . Figs. 1-4. Chrysosphaerella brevispina. Bacteria present in the perinuclear space (arrows). Figure 4 illustrates a dividing cell with two nuclei. Scale = 1 m. non-infected cells. Bacteria numbered from one to eight, with the usual number ranging between two to three. Bacteria were not observed in the cytoplasm of the cell. If bacteria were present in cytoplasmic vesicles as ingested particles, they could not be disThis content downloaded from 157.55.39.212 on Thu, 09 Jun 2016 06:24:57 UTC All use subject to http://about.jstor.org/terms 34 TRANSACTIONS OF THE KANSAS ACADEMY OF SCIENCE tinguished. The bacteria present in the perinuclear space showed no signs of being digested and appeared similar in gross morphological features to bacteria observed as endosymbionts in other algae. The presence of bacteria between the cell membrane and siliceous scale covering was not observed. Similar observations were made in a related genus, Mallomonas (Belcher, 1969); however, in a different species of Mallomonas, bacteria were shown to be living in this area (Wujek, 1978). Attempts to observe the bacteria with light microscopy using a variety of bacterial stains were not productive. These attempts included both phase and Nomarski contrast. The transmission of the bacteria from one cell to another appears to take place during cell division (Fig. 4). When the nucleus divides, one or more bacteria move with the daughter nuclei into the new daughter cells. The algae showed no external or internal damage from the bacteria (aside from the swollen perinuclear space). As cells within the same colony were observed with no bacteria, what benefit, if any, was gained from the association was not determined. Because cysts were not observed, it could not be determined if the statospore harbors bacteria during overwintering.
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