Supporting Online Material J Bacteriol

1Supporting online material J Bacteriol

2Movie S1

3Bdellovibrio growth-phase cells septate synchronously in bdelloplasts. Time-lapse movie

4showing a bdelloplast of an infected fluorescent prey cell with a dark growing Bdellovibrio cell

5within; this cell elongates from both poles until it reaches its maximum length and septates

6synchronously into 5 progeny, which lyse the bdelloplast resulting in the loss of fluorescence

7activity. Contrast shifts within the bdelloplast are likely due to shifts in the prey cytoplasmic

8membrane. Images were captured every 2.5 min and shown at 7 frames per second, scale bar = 1

10Movie S2

11Two separate Bdellovibrio growth-phase cells synchronously septate in doubly infected

12bdelloplasts. Time-lapse movie showing a bdelloplast of an infected fluorescent prey cell with

13two separate dark growing Bdellovibrio cells within; these cells elongate from both poles, until

14they reach their maximum length and septate synchronously into a total of 7 progeny (4 from the

15lower and 3 from the upper cell), which lyse the bdelloplast resulting in the loss of fluorescence

16activity. Growing cells seem to favour the bdelloplast periphery, presumably due to an unseen

17obstacle (for example the prey cell cytoplasm) in the centre; this inhibition seems to be released

18in the final stages of septation. Images were captured every 2.5 min and shown at 7 frames per

19second, scale bar = 1 µm.

20Movie S3

21Long Bdellovibrio growth-phase cells initiate septation synchronously in bdelloplasts. Time-

22lapse movie showing a bdelloplast of an infected fluorescent prey cell with a dark growing

23Bdellovibrio cell within; this cell elongates from both poles until it reaches its maximum length

2 24and initiates septation synchronously along an extended filament which will ultimately generate

258 progeny. Right hand pole of the elongating Bdellovibrio cell seems to get caught on an unseen

26obstacle, which is released in the later stages of growth. Contrast shifts within the bdelloplast are

27likely due to shifts in the prey cytoplasmic membrane. Growing cells seem to favour the

28bdelloplast periphery, presumably due to an unseen obstacle (for example the prey cell

29cytoplasm) in the centre; this inhibition seems to be released in the final stages of cell elongation.

30Images were captured every 2.5 min and shown at 7 frames per second, scale bar = 1 µm.

31Movie S4

32Stored tension within a Bdellovibrio growth-phase filament leads to dramatic re-orientation of

33progeny cells after synchronous septation. Time-lapse movie, showing a bdelloplast of an

34infected fluorescent prey cell with a dark elongated and coiled Bdellovibrio cell within; this

35growth-phase cell septates leading to a dramatic shift in orientation of the 5 progeny cells, only

36possible if septation was synchronous. Progeny cells lyse the bdelloplast resulting in the loss of

37fluorescence activity. Images were captured every 2.5 min and shown at 7 frames per second,

38scale bar = 1 µm.

41Movie S5

42Bdellovibrio enter prey cells and, some, rarely, do not elongate and divide to form new progeny

43cells. Time-lapse dual-channel movie of overlaid brightfield (red) and fluorescence (green)

44images, initial fluorescence bleaching shows an example of a single dark Bdellovibrio cell in a

45small, fluorescent, rounded bdelloplast. The Bdellovibrio cell does not elongate or septate, and

46persists in the bdelloplast eventually lysing it (leading to a loss of fluorescence activity) and the

4 47single cell is clearly seen escaping to the lower right of the frame. Images were captured every

482.5 min and shown at 7 frames per second, scale bar = 1 µm.

50Movie S6

51Mature Bdellovibrio progeny escape the bdelloplast through a single hole. Time-lapse movies of

52overlaid brightfield (red) and fluorescence (green) images, movie shows a bdelloplast of an

53infected fluorescent prey cell with a dark growing Bdellovibrio cell within, and is the same

54bdelloplast shown in Movie S1. The growing Bdellovibrio cell septates synchronously into 5

55progeny, which lyse the bdelloplast resulting in loss of fluorescence activity; progeny cells are

56then seen leaving the now exhausted prey cell through a hole in the bottom of the bdelloplast.

57This pattern was seen in 88% of observed bdelloplasts (n=67). Images were captured every 2.5

58min and shown at 7 frames per second, scale bar = 1 µm.

59Movie S7

60Time-lapse brightfield movie showing motile Bdellovibrio progeny cells within a bdelloplast, at

61maturation, after septation. Three Bdellovibrio progeny cells appear to scramble over one another

62and collide with the inner wall of the bdelloplast which is immobilised on a glass surface in

63Ca/HEPES buffer. Internal collisions occasionally distort the shape of the bdelloplast. Images

64were captured at 25 frames per second using the IPLab software (version 3.64) and are shown at

65the same rate, scale bar = 1 µm.