(4) Several of these species of Coleoptera have a synizesis stage in which the spermatogonial number of short loops is massed at one side of the nucleus. This is followed by a synapsis stage in which the loops straighten and unite in pairs, forming longer loops which soon spread out in the nuclear space, and, with the exception of the heterochromosomes, unite to form a continuous spireme.
(5) In several of the species of Coleoptera and in Aphrophora, it has been shown that a body staining like chromatin develops in the spermatids, increasing in size for a time, then breaking up into granules and disappearing. This body evidently has no relation to the heterochromosomes, as it is the same for all of the spermatids. Its staining qualities suggest that it may be material derived from the chromosomes. It is finally dissolved in the karyolymph.
(6) In iron-hæmatoxylin preparations the heterochromosomes of the Coleoptera vary greatly in their staining properties during mitosis. In some species they stain exactly like the ordinary chromosomes, in others the larger one of the unequal pair holds the stain more tenaciously than the others and also than its smaller mate, and this is true in several cases where the heterochromosome is smaller than the other chromosomes, which destain more readily. The odd chromosome of the Elaters stains less deeply than the others in the first spermatocyte. In the growth stage they stain more deeply, as a rule, than the spireme, with iron-hæmatoxylin or thionin, stain red with safranin-gentian and green with Auerbach's methyl green-fuchsin combination.
(7) Aphrophora quadrangularis agrees with the Anasa group of Hemiptera heteroptera in having a pair of m-chromosomes and an odd chromosome in the spermatocytes, but differs from many of that group in that the odd chromosome divides in the second mitosis instead of the first. It also differs from other known forms in exhibiting heterochromosomes in certain stages of the oöcytes.
(8) The two species of Lepidoptera examined have an equal pair of heterochromosomes.
FOOTNOTES:
[A] Aug. 20, 1906.—Since this paper was prepared, 19 other species of Coleoptera have been studied. Of these, 17 have an unequal pair of heterochromosomes in the spermatocytes. Six belong to the Chrysomelidæ, making 14 of that family that have been examined. Representatives of 4 new families—Melandryidæ, Lamiinæ, Meloidæ, Cerambycinæ have been studied. In only two species—1 Elater and 1 Lampyrid—has the odd chromosome been found in place of the unequal pair. No species of Coleoptera has yet been examined in which one or the other of these two types of heterochromosomes does not occur in the spermatocytes. Of the 42 species of Coleoptera whose germ cells have been studied, 85.7 per cent are characterized by the presence of an unequal pair of heterochromosomes in the male germ cells, 14.3 per cent by the presence of an odd chromosome.