Werren John H, Loehlin David W
Department of Biology, University of Rochester, Rochester, NY 14627, USA.
Cold Spring Harb Protoc. 2009 Oct;2009(10):pdb.prot5310. doi: 10.1101/pdb.prot5310.
This protocol covers most procedures used by the Nasonia (wasp) geneticist, including virgin collection. Nasonia are easily sexed as pupae, so collecting virgins can easily be performed within a window of several days. The female-specific ovipositor is visible at even the earliest pupal stages as a longitudinal ridge on the ventral posterior abdomen. To prolong generations, yellow pupae can be stored at 4 degrees C-8 degrees C for up to 2 mo and adults for 2-3 wk. In the example presented here, haplodiploid crossing methods are used to make a new Nasonia vitripennis mutant homozygous, using an aunt/nephew mating approach. The protocol can be extended to inbreeding strains. Inbreeding depression generally is not a problem in Nasonia unless the mutation itself is female-sterile. Mother/son mating and sib mating are also reasonable approaches to inbreeding.
本方案涵盖了纳氏胡蜂（黄蜂）遗传学家使用的大多数程序，包括处女蜂采集。纳氏胡蜂在蛹期很容易区分性别，所以在几天的时间窗口内就可以轻松地采集到处女蜂。即使在最早的蛹期，雌性特有的产卵器也可见，表现为腹侧后腹部的一条纵向脊。为了延长世代，可以将黄色蛹在4摄氏度至8摄氏度下储存长达2个月，成虫可储存2至3周。在此处给出的示例中，使用单双倍体杂交方法，采用姑侄交配方式使新的丽蝇蛹集金小蜂突变体纯合。该方案可扩展到近交系。除非突变本身是雌性不育的，否则近交衰退在纳氏胡蜂中一般不是问题。母子交配和同胞交配也是近交的合理方法。
