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拟步甲属(鞘翅目:金花虫科)色氨酸 2,3-双加氧酶基因的特征及 RNA 干扰介导的沉默。

Characterization and RNA interference-mediated silencing of tryptophan 2,3-dioxygenase gene in Carpophilus hemipterus (L.) (Coleoptera: Nitidulidae).

机构信息

USDA-ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, California, USA.

USDA-ARS, National Soil Dynamics Research Laboratory, Auburn, Alabama, USA.

出版信息

Arch Insect Biochem Physiol. 2024 Jan;115(1):e22072. doi: 10.1002/arch.22072.

Abstract

Dried fruit beetle, Carpophilus hemipterus (Linnaeus, 1758) (Coleoptera: Nitidulidae), is a serious pest of ripened fresh fruit in the orchard and dried fruit in postprocessing storage. Despite the economic impact and widespread distribution of C. hemipterus, there is a lack of functional genomics research seeking to elucidate features of molecular physiology for improved pest management. Here, we report the characterization of the gene named Vermilion in C. hemipterus (ChVer) that encodes for tryptophan 2,3-dioxygenase. The Vermilion is frequently used as a visual marker for genomics approaches as tryptophan 2,3-dioxygenase is involved in the biosynthesis of eye coloration pigments in insects. We identified 1628 bp long full-length transcript of ChVer from transcriptomic database of C. hemipterus. The expression analysis among adult body parts revealed peak ChVer expression in head compared to thorax and abdomen, which is consistent with its role. Among the C. hemipterus developmental stages, peak ChVer expression was observed in first instar larva, second instar larva, and adult male stages, whereas the lowest levels of expression were seen in third instar larva, prepupa, and pupa. The nanoinjection of ChVer double-stranded RNA in larval C. hemipterus resulted in a significant reduction in ChVer transcript levels as well as caused a loss of eye color, that is, the white-eyed phenotype in adults. Characterization of visually traceable marker gene and robust RNA interference response seen in this study will enable genomics research is this important pest.

摘要

干果甲虫, Carpophilus hemipterus(Linnaeus,1758)(鞘翅目:丽金龟科),是果园中成熟新鲜水果和加工后储存的干果的严重害虫。尽管 C. hemipterus 具有经济影响且分布广泛,但缺乏功能基因组学研究,无法阐明其分子生理学特征,以实现更好的害虫管理。在这里,我们报告了 C. hemipterus 中名为 Vermilion 的基因(ChVer)的特征,该基因编码色氨酸 2,3-双加氧酶。Vermilion 经常被用作基因组学方法的视觉标记,因为色氨酸 2,3-双加氧酶参与昆虫眼色色素的生物合成。我们从 C. hemipterus 的转录组数据库中鉴定出长 1628bp 的全长 ChVer 转录本。在成虫身体部位的表达分析中,与胸部和腹部相比,头部的 ChVer 表达量最高,这与其作用一致。在 C. hemipterus 的发育阶段中,在第一龄幼虫、第二龄幼虫和雄性成虫阶段观察到 ChVer 表达峰值,而在第三龄幼虫、预蛹和蛹阶段表达水平最低。在幼虫 C. hemipterus 中注射 ChVer 双链 RNA 会导致 ChVer 转录本水平显著降低,并导致眼睛颜色丧失,即成虫出现白眼表型。本研究中观察到可追踪的视觉标记基因的特征和强大的 RNA 干扰反应,将使该重要害虫的基因组学研究成为可能。

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