Brent Colin S, Heu Chan C, Gross Roni J, Fan Baochan, Langhorst Daniel, Hull J Joe
USDA-ARS Arid Land Agricultural Research Center, Maricopa, AZ 85138, USA.
Insects. 2022 Oct 27;13(11):986. doi: 10.3390/insects13110986.
Cuticle coloration in insects is a consequence of the accumulation of pigments in a species-specific pattern. Numerous genes are involved in regulating the underlying processes of melanization and sclerotization, and their manipulation can be used to create externally visible markers of successful gene editing. To clarify the roles for many of these genes and examine their suitability as phenotypic markers in Knight (western tarnished plant bug), transcriptomic data were screened for sequences exhibiting homology with the proteins. Complete open reading frames encoding putative homologs for six genes (, , , , , and ) were identified, with two variants for . Sequence and phylogenetic analyses supported preliminary annotations as cuticle pigmentation genes. In accord with observable difference in color patterning, expression varied for each gene by developmental stage, adult age, body part, and sex. Knockdown by injection of dsRNA for each gene produced varied effects in adults, ranging from the non-detectable (, ), to moderate decreases (, ) and increases (, ) in darkness, to extreme melanization (). Based solely on its expression profile and highly visible phenotype, appears to be the best marker for tracking transgenic .
昆虫表皮着色是色素以物种特异性模式积累的结果。许多基因参与调节黑化和硬化的潜在过程，对这些基因的操控可用于创建成功基因编辑的外部可见标记。为了阐明这些基因中许多基因的作用，并检验它们作为克奈特氏虫（西部变色蝽象）表型标记的适用性，对转录组数据进行筛选，以寻找与这些蛋白质具有同源性的序列。鉴定出了编码六个基因（、、、、和）假定同源物的完整开放阅读框，其中有两个变体。序列和系统发育分析支持将其初步注释为表皮色素沉着基因。与观察到的颜色图案差异一致，每个基因的表达随发育阶段、成虫年龄、身体部位和性别而变化。通过注射针对每个基因的dsRNA进行基因敲低，在成虫中产生了不同的效果，从不可检测（、）到黑暗中适度减少（、）和增加（、），再到极端黑化（）。仅基于其表达谱和高度可见的表型，似乎是追踪转基因的最佳标记。
