Perkins D D
Genetics. 1974 Jul;77(3):459-89. doi: 10.1093/genetics/77.3.459.
Rapid, effective techniques have been developed for detecting and characterizing chromosome aberrations in Neurospora by visual inspection of ascospores and asci. Rearrangements that are detectable by the presence of deficient, nonblack ascospores in test crosses make up 5 to 10% of survivors after UV doses giving 10-55% survival. Over 135 rearrangements have been diagnosed by classifying unordered asci according to numbers of defective spores. (These include 15 originally identified or analyzed by other workers.) About 100 reciprocal translocations (RT's) have been confirmed and mapped genetically, involving all combinations of the seven chromosomes. Thirty-three other rearrangements generate viable nontandem duplications in meiosis. These consist of insertional translocations (IT's) (15 confirmed), and of rearrangements that involve a chromosome tip (10 translocations and 3 pericentric inversions). No inversion has been found that does not include the centromere. A reciprocal translocation was found within one population in nature. When pairs of RT's that involve the same two chromosome arms were intercrossed, viable duplications were produced if the breakpoints overlapped in such a way that pairing resembled that of insertional translocations (27 combinations).-The rapid analytical technique depends on the following. Deficiency ascospores are usually nonblack (W: "white") and inviable, while nondeficient ascospores, even those that include duplications, are black (B) and viable. Thus RT's typically produce 50% black spores, and IT's 75% black. Asci are shot spontaneously from ripe perithecia, and can be collected in large numbers as groups of eight ascospores representing unordered tetrads, which fall into five classes: 8B:0W; 6B:2W, 4B:4W, 2B:6B, 0B:8W. In isosequential crosses, 90-95% of tetrads are 8:0. When a rearrangement is heterozygous, the frequencies of tetrad classes are diagnostic of the type of rearrangement, and provide information also on the positions of break points. With RT's, 8:0 (alternate centromere segregation) = 0:8 (adjacent-1), 4:4's require interstitial crossing over in a centromere-break point interval, and no 6:2's or 2:6's are expected. With IT's, duplications are viable, 8:0 = 4:4, 6:2's are from interstitial crossing over, 0:8's or 2:6's are rare. Tetrads from RT's that involve a chromosome tip resemble those from IT's, as do tetrads from intercrosses between partially overlapping RT's that involve identical chromosome arms.-Because viable duplications and other aneuploid derivatives regularly occur among the offspring of rearrangements such as insertional translocations, care must be taken in selecting stocks, and original strains should be kept for reference.
通过对视察子囊孢子和子囊来检测和鉴定粗糙脉孢菌染色体畸变,已经开发出快速、有效的技术。在测试杂交中,通过存在缺失、非黑色子囊孢子可检测到的重排占紫外线剂量后存活者的5%至10%,该紫外线剂量使存活率为10 - 55%。通过根据有缺陷孢子的数量对无序子囊进行分类,已诊断出超过135种重排。(其中包括其他研究人员最初鉴定或分析的15种。)大约100种相互易位(RT)已得到确认并进行了遗传定位,涉及七条染色体的所有组合。另外33种重排在减数分裂中产生可行的非串联重复。这些包括插入易位(IT)(15种已确认),以及涉及染色体末端的重排(10种易位和3种臂间倒位)。未发现不包括着丝粒的倒位。在自然界的一个群体中发现了一种相互易位。当涉及相同两条染色体臂的RT对相互杂交时,如果断点以类似于插入易位的配对方式重叠,则会产生可行的重复(27种组合)。 - 这种快速分析技术依赖于以下几点。缺失子囊孢子通常是非黑色(W:“白色”)且不可存活,而非缺失子囊孢子,即使那些包括重复的,也是黑色(B)且可存活。因此,RT通常产生50%的黑色孢子,IT产生75%的黑色孢子。子囊从成熟的子囊中自发射出,可以大量收集作为代表无序四分体的八个子囊孢子组,它们分为五类:8B:0W;6B:2W、4B:4W、2B:6W、0B:8W。在等序列杂交中,90 - 95%的四分体是8:0。当重排是杂合时,四分体类别的频率可诊断重排类型,并还提供有关断点位置的信息。对于RT,8:0(交替着丝粒分离) = 0:8(相邻 - 1),4:4需要在着丝粒 - 断点区间进行中间交换,并且预计不会出现6: