Filling in the gaps: A reevaluation of the Lygus hesperus peptidome using an expanded de novo assembled transcriptome and molecular cloning
Filling in the gaps: A reevaluation of the Lygus hesperus peptidome using an expanded de novo assembled transcriptome and molecular cloning
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DOI：
10.1016/j.ygcen.2020.113708
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发表时间：
2021-01-13
影响因子：
2.7
通讯作者：
Christie, Andrew E.
中科院分区：
文献类型：
--
作者：
Hull, J. Joe;Gross, Roni J.;Christie, Andrew E.
肽是调节生理和行为的最大和最多样化的一类分子。在此之前，我们预测了一个peptidome的西方失去光泽的植物错误，草盲蝽，使用从整个个人的转录组数据。该分析的一个潜在限制是掩盖了代表性不足的基因，特别是组织特异性转录本。在这里，我们重新评估了L。hesperus peptidome使用更全面的数据集，该数据集包括先前的转录组学数据以及从头部和附属腺产生的组织特异性读数。这种增强的组装显著提高了覆盖深度，提供了基本上所有先前鉴定的家族的确证性转录物和编码对应于14个肽家族的许多新肽前体的新转录物。在我们最初的研究中没有针对的几个家族在扩展的组装中被鉴定，包括agatoxin样肽、CNMamide、神经肽样前体1和periviscerokinin。为了增加计算机模拟数据的置信度，使用RT-PCR扩增新鉴定的转录物子集的开放阅读框并进行序列验证。进一步基于PCR的分析推定的L. hesperus agatoxin样肽前体揭示了在L. hesperus的发展，表明肽的功能作用超出了毒素。这里预测的肽，结合我们早期研究中鉴定的肽，扩展了L。hesperus peptidome已扩展到42个家族成员，并为启动对这种非模式农业害虫的肽能功能的分子和生理研究提供了一个改进的平台。
Peptides are the largest and most diverse class of molecules modulating physiology and behavior. Previously, we predicted a peptidome for the western tarnished plant bug, Lygus hesperus, using transcriptomic data produced from whole individuals. A potential limitation of that analysis was the masking of underrepresented genes, in particular tissue-specific transcripts. Here, we reassessed the L. hesperus peptidome using a more comprehensive dataset comprised of the previous transcriptomic data as well as tissue-specific reads produced from heads and accessory glands. This augmented assembly significantly improves coverage depth providing confirmatory transcripts for essentially all of the previously identified families and new transcripts encoding a number of new peptide precursors corresponding to 14 peptide families. Several families not targeted in our initial study were identified in the expanded assembly, including agatoxin-like peptide, CNMamide, neuropeptide-like precursor 1, and periviscerokinin. To increase confidence in the in silico data, open reading frames of a subset of the newly identified transcripts were amplified using RT-PCR and sequence validated. Further PCR-based profiling of the putative L. hesperus agatoxin-like peptide precursor revealed evidence of alternative splicing with near ubiquitous expression across L. hesperus development, suggesting the peptide serves functional roles beyond that of a toxin. The peptides predicted here, in combination with those identified in our earlier study, expand the L. hesperus peptidome to 42 family members and provide an improved platform for initiating molecular and physiological investigations into peptidergic functionality in this non-model agricultural pest.