Use of Neurospora Spore killer strains to obtain centromere linkage data without dissecting asci

Abstract

Use of a centromere-linked Spore killer gene Sk reduces manyfold the labor involved in obtaining tetrad data that would otherwise require ordered dissection of intact linear eight-spored asci. Heterozygous crosses are made for Spore killer (Sk^K × Sk^S) and for markers to be tested. In such crosses only Sk^K ascospores survive. The four viable (Sk^K) and four aborted (Sk^S) ascospores of each ascus are ejected from the perithecium as a physically disordered group. The four surviving Sk^K ascospores of individual asci are germinated and scored. Sk^K segregates from Sk^S at the first meiotic division. If both marker alleles are represented in the surviving products, they must therefore have segregated from one another at the second division. Four-spore (Fsp) genes have been used to eliminate one postmeiotic nuclear division, so that only two ascospores per ascus need to be scored. The Spore killer method has been useful for mapping closely linked genes in centromere regions, for identifying genes that are far out on chromosome arms, for obtaining information on meiotic crossing-over, and for comparing linkages in different species.Key words: tetrad analysis, centromere mapping, Spore killer, Neurospora.