Tabel of Pupal Stages

Stages of metamorphosis in Drosophila melanogaster (at 25°C)
Stage	Time (hours)^a	Developmental event
P1	0–1	White puparium: wriggling stops completely
P2	1–3	Brown puparium: oral armature stops moving permanently, heart stops pumping, gas bubble becomes visible within abdomen
P3	3–6.5	Bubble prepupa: puparium becomes separated from underlying epidermis; bubble in abdominal region is large, causing prepupa to become positively buoyant at end of this stage (it floats)
P4	6.5–12.5	Buoyant and moving bubble: prepupa is buoyant, and bubble moves, first appearing in the posterior of the puparium, displacing pupa anteriorly, and then appearing in the anterior, displacing the pupa posteriorly. Imaginal head sac is everted and oral armature of larva is expelled
P5	12.5–25	Malpighian tubules migrating and white: legs and wings extend; Malpighian tubules move from thorax to abdomen and become visible as white structures in dorsal anterior abdomen
P6	25–43	Green Malpighian tubules: Malpighian tubules turn green, and dark green “yellow body” appears between the anterior ends of the two Malpighian tubules
P7	43–47	“Yellow body”: “yellow body” (actually dark green) moves back between the Malpighian tubules; transparent pupal cuticle separates from underlying epidermis; eye cup becomes yellow at its perimeter
P8	47–57	Yellow-eyed: eyes become bright yellow
P9	57–69	Amber: eyes darken to deep amber
P10	69–73	Red-eye Bald: eyes become bright red; orbital and ocellar bristles and vibrissae darken
P11	73–78	Head and thoracic bristles: head bristles, followed by thoracic bristles, darken
P12	73–78	Wings grey: wings become gray; sex comb darkens
P13	78–87	Wings black: wings become black; tarsal bristles darken and claws become black
14	87–90	Mature bristles: green patch (the meconium–waste products of pupal metabolism) appears dorsally at posterior tip of abdomen
P15	90–103	Meconium and eclosion: tergites become tan, obscuring Malpighian tubules and “yellow body”; legs twitch; flies able to walk prematurely if puparium removed; eclosion completed
^aTimes start on approximately day four of larval life when the larva is still white but is no longer able to crawl. Timing is variable among individuals, and the times given are a simplification from the Bainbridge and Bownes paper. Source: After Bainbridge, S. P. and M. Bownes. 1981. Staging the metamorphosis of Drosophila melanogaster. J. Embryol. Exp. Morph. 66: 57–80.