Megachile kyotensis Alfken is a small leaf-cutting bee species endemic to Japan, with a univoltine life cycle occurring from mid July to mid September. Its nesting biology and nest architecture were studied in Nasukarasuyama, Tochigi Pref., central Japan from 2009 to 2011. This is the first authentic record of the nesting biology for this species. Nest burrows were excavated by the bee herself in brownish, loamy soil of the gentle slope. The burrows were 7-8 mm in diameter and 5-7 cm long from the entrance to the bottom of the deepest brood cell. In completed nests 3-6 branch burrows diverged from the main burrows. Entrance closure and burrow filling with leaf pieces were sporadically observed in the completed nests. The number of brood cells per branch burrow differed among nests. The number of brood cells per nest was 4-10 (mean 6.0), which was relatively large among soil-burrowing, leaf-cutting bees. Several innermost leaf pieces of both cell cups and cell caps were firmly stuck together using some adhesive substance. Completed cells were made of five different types of leaf pieces: A-1 large, oval-shaped; A-2 small, oval-shaped; A-3 small, oval-shaped, used for cell bottom; B-1 round-shaped; and B-2 semi-round shaped. A-type pieces were used for cell cups and B-type ones for cell caps. These leaf pieces were very small, probably corresponding to the body size of this species. The oval-shaped pieces were relatively uniform in size, 85% ranging from 12.1 to 15.0 mm in length. The mean number of leaf pieces per cell was 17.4 (10.8 for the cup and 6.6 for the cap), which was much less than those in other soilburrowing, leaf-cutting bees except M. rixator sakishimana Yasumatsu et Hirashima. In M. kyotensis all leaf pieces were used so as that the upper side of leaf was directed toward the inside of brood cell. On the average, the cocoon size was 10.5×5.6 mm in females and 9.8×5.5 mm in males, and the cocoon wall was 0.11 mm in thickness. A cleptoparasitic fly, Miltogramma angustifrons (Townsend) (Sarcophagidae), and a cleptoparasitic blister beetle, Zonitis japonica Pic (Meloidae), were recorded from the nests of M. kyotensis. The former was an important enemy, attacking nine out of 30 brood cells.
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