Ghanim Murad, Dombrovsky Aviv, Raccah Benny, Sherman Amir
Department of Entomology, The Volcani Center, P.O. Box 6 Bet Dagan 50250, Israel.
Insect Biochem Mol Biol. 2006 Nov;36(11):857-68. doi: 10.1016/j.ibmb.2006.08.007. Epub 2006 Sep 3.
Aphids exhibit a complex life cycle in which their phenotypes vary in response to environmental changes. Wing polyphenism is one of the phenotypes that have not been adequately studied. We developed a cDNA microarray from the green peach aphid Myzus persicae (Sulzer), and compared mRNA samples derived from alate and apterous adults. The microarray experiment resulted in 108 significantly changing clones that represented 31 unique ESTs. Among the highest and significantly expressed clones in alate adults, seven clones showed portions of a new adenine nucleotide translocase (ANT) gene from M. persicae and four clones contained sequences of the OS-D gene. Another clone showed significant homology with the takeout (TO) and TO-like proteins described from other insects. A full-length cDNA clone of M. persicae ANT (Mp ANT) was isolated and characterized. An reverse transcription-polymerase chain reaction (RT-PCR) analysis showed significant differences in Mp ANT, OS-D and Mp takeout-like genes (Mp TOL) expression in alate compared to apterous adults. A polyclonal antibody against the beef heart mitochondrial ANT reacted with a recombinant Mp ANT expressed in Escherichia coli, as it did with aphid extracts, showing higher amounts of the expected 33kDa band in alate adults. Spatial expression analysis showed higher Mp ANT expression in the thorax compared to head, abdomen, leg and antennae. OS-D and Mp TOL showed ubiquitous expression across the body, with OS-D being significantly higher in legs and antennae while Mp TOL being significantly higher in the head and abdomen. The possible causes for higher expression of ANT, OS-D and TOL in alates and the physiological effects on the aphid flight and dispersal are discussed.
蚜虫表现出复杂的生命周期,其表型会随环境变化而改变。翅多型现象是其中一种尚未得到充分研究的表型。我们构建了来自桃蚜(烟蚜茧蜂)的cDNA微阵列,并比较了有翅和无翅成虫的mRNA样本。微阵列实验产生了108个显著变化的克隆,代表31个独特的EST。在有翅成虫中高表达且显著的克隆中,7个克隆显示了来自桃蚜的新腺嘌呤核苷酸转位酶(ANT)基因的部分序列,4个克隆包含OS-D基因的序列。另一个克隆与其他昆虫中描述的外卖(TO)和TO样蛋白具有显著同源性。分离并鉴定了桃蚜ANT(Mp ANT)的全长cDNA克隆。逆转录-聚合酶链反应(RT-PCR)分析表明,与无翅成虫相比,有翅成虫中Mp ANT、OS-D和Mp外卖样基因(Mp TOL)的表达存在显著差异。针对牛心线粒体ANT的多克隆抗体与在大肠杆菌中表达的重组Mp ANT反应,就像与蚜虫提取物反应一样,在有翅成虫中显示出预期的33kDa条带含量更高。空间表达分析表明,与头部、腹部、腿部和触角相比,胸部的Mp ANT表达更高。OS-D和Mp TOL在全身普遍表达,OS-D在腿部和触角中显著更高,而Mp TOL在头部和腹部中显著更高。讨论了有翅成虫中ANT、OS-D和TOL表达较高的可能原因以及对蚜虫飞行和扩散的生理影响。