Walters Michelle, Morrison Neil I, Claus John, Tang Guolei, Phillips Caroline E, Young Robin, Zink Richard T, Alphey Luke
Animal Plant Health and Inspection Service, Centers for Plant Health Science and Technology, United States Department of Agriculture, Phoenix, Arizona, United States of America.
PLoS One. 2012;7(6):e38547. doi: 10.1371/journal.pone.0038547. Epub 2012 Jun 5.
The cotton pest, pink bollworm (Pectinophora gossypiella (Saunders)), is a significant pest in most cotton-growing areas around the world. In southwestern USA and northern Mexico, pink bollworm is the target of the sterile insect technique (SIT), which relies on the mass-release of sterile pink bollworm adults to over-flood the wild population and thereby reduce it over time. Sterile moths reared for release are currently marked with a dye provided in their larval diet. There are concerns, however, that this marker fails from time to time, leading to sterile moths being misidentified in monitoring traps as wild moths. This can lead to expensive reactionary releases of sterile moths. We have developed a genetically marked strain that is engineered to express a fluorescent protein, DsRed2, which is easily screened under a specialised microscope. In order to test this marker under field conditions, we placed wild-type and genetically marked moths on traps and placed them in field cages. The moths were then screened, in a double-blind fashion, for DsRed2 fluorescence at regular intervals to determine marker reliability over time. The marker was shown to be robust in very high temperatures and generally proved reliable for a week or longer. More importantly, genotyping of moths on traps by PCR screening of the moths was 100% correct. Our findings indicate that this strain--and fluorescent protein markers in general--could make a valuable contribution to SIT.
棉铃虫(Pectinophora gossypiella (Saunders))是全球大多数棉花种植区的一种重要害虫。在美国西南部和墨西哥北部,棉铃虫是昆虫不育技术(SIT)的防治对象,该技术依靠大量释放不育的棉铃虫成虫来淹没野生种群,从而随着时间的推移减少其数量。目前,用于释放的不育蛾是通过在其幼虫饲料中添加染料来标记的。然而,人们担心这种标记有时会失效,导致不育蛾在监测诱捕器中被误认成野生蛾。这可能会导致昂贵的不育蛾应急释放。我们开发了一种基因标记品系,通过基因工程使其表达一种荧光蛋白DsRed2,这种蛋白在专门的显微镜下易于筛选。为了在田间条件下测试这种标记,我们将野生型和基因标记的蛾子放置在诱捕器上,并将它们放置在田间笼子里。然后,以双盲方式定期对蛾子进行DsRed2荧光筛选,以确定标记随时间的可靠性。结果表明,该标记在非常高的温度下依然稳定,并且在一周或更长时间内总体上证明是可靠的。更重要的是,通过对诱捕器上的蛾子进行PCR筛选进行基因分型的结果100%正确。我们的研究结果表明,这种品系——以及一般的荧光蛋白标记——可以为昆虫不育技术做出有价值的贡献。