Odoriferous Defensive stink gland transcriptome to identify novel genes necessary for quinone synthesis in the red flour beetle, Tribolium castaneum.
Odoriferous Defensive stink gland transcriptome to identify novel genes necessary for quinone synthesis in the red flour beetle, Tribolium castaneum.
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DOI：
10.1371/journal.pgen.1003596
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发表时间：
2013
期刊：
影响因子：
4.5
通讯作者：
Wimmer EA
中科院分区：
文献类型：
--
作者：
Li J;Lehmann S;Weißbecker B;Ojeda Naharros I;Schütz S;Joop G;Wimmer EA
化学防御是昆虫最重要的特征之一，它赋予昆虫征服最多样化的生态环境的能力。化学分泌物被用来防御从脊椎动物或无脊椎动物捕食者到原核生物或真核生物寄生虫或食物竞争对手的任何东西。天牛在这一类中尤其多产，能产生几种取代的苯醌化合物。为了更好地了解防御性分泌物的遗传和分子基础，我们对一种新出现的昆虫模型--赤粉甲虫进行了RNA测序。为了检测在分泌防御性化合物的恶臭腺体组织中高度和特异表达的基因，我们将它们与对照组织前腹部进行了比较。在不同的消减组中共鉴定出511个基因。其中，77个基因通过RNA干扰(RNAi)进行了功能分析，以识别诱导的腺体形态变化或通过气相色谱-质谱法识别腺体挥发物的变化。29个基因(38%)表现出强烈的可见表型，而67个基因(87%)表现出至少一个腺体内容物的变化。分离并鉴定了其中三个具有无醌(Ql)表型的基因-TCAS-ql VTG1；TCAS-ql ARSB；TCAS-ql MRP，鉴定了它们在两种类型的分泌腺细胞中的表达，并通过RNAi后所有主要成分的定量来确定它们的功能。此外，微生物抑制试验表明，无醌状态不能阻碍细菌或真菌的生长。对这三个基因的系统发育分析表明，它们是为甲虫的化学防御而独立进化的。昆虫使用化学防御性分泌物来保护自己免受周围所有潜在的有害因素的伤害。天牛产生几种取代的苯醌化合物作为刺激物和驱避剂，这些化合物具有高度的活性和毒性。然而，受控合成和分泌的分子基础仍有待鉴定。在这项研究中，我们使用了一个新出现的昆虫模型--赤粉甲虫，通过鉴定分泌腺中高表达和特异表达的基因，更好地了解了苯二酚产生的遗传和分子基础。通过对不同的腺体组织进行RNA测序和转录，我们鉴定了511个腺体特异基因。其中，77个基因被功能检测腺体形态变化和腺体挥发性成分的变化。我们发现，在基于RNA干扰的基因敲除后，67个基因(87%)导致至少一个腺体挥发性改变。对三个新基因进行了进一步鉴定，以证实它们在防御性分泌物中的重要性及其独立的进化起源。
Chemical defense is one of the most important traits, which endow insects the ability to conquer a most diverse set of ecological environments. Chemical secretions are used for defense against anything from vertebrate or invertebrate predators to prokaryotic or eukaryotic parasites or food competitors. Tenebrionid beetles are especially prolific in this category, producing several varieties of substituted benzoquinone compounds. In order to get a better understanding of the genetic and molecular basis of defensive secretions, we performed RNA sequencing in a newly emerging insect model, the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae). To detect genes that are highly and specifically expressed in the odoriferous gland tissues that secret defensive chemical compounds, we compared them to a control tissue, the anterior abdomen. 511 genes were identified in different subtraction groups. Of these, 77 genes were functionally analyzed by RNA interference (RNAi) to recognize induced gland alterations morphologically or changes in gland volatiles by gas chromatography-mass spectrometry. 29 genes (38%) presented strong visible phenotypes, while 67 genes (87%) showed alterations of at least one gland content. Three of these genes showing quinone-less (ql) phenotypes – Tcas-ql VTGl; Tcas-ql ARSB; Tcas-ql MRP – were isolated, molecularly characterized, their expression identified in both types of the secretory glandular cells, and their function determined by quantification of all main components after RNAi. In addition, microbe inhibition assays revealed that a quinone-free status is unable to impede bacterial or fungal growth. Phylogenetic analyses of these three genes indicate that they have evolved independently and specifically for chemical defense in beetles. Insects use chemical defensive secretions to defend themselves from all the potential detrimental factors in their surroundings. Tenebrionid beetles produce as irritants and repellents several varieties of substituted benzoquinone compounds that are highly reactive and toxic. However, the molecular basis of the controlled synthesis and secretion remains to be identified. In this study, we employ a newly emerging insect model, the red flour beetle Tribolium castaneum, to get a better understanding of the genetic and molecular basis of quinone production with identification of the highly and specifically expressed genes in secretory glands. By performing RNA sequencing and transcriptomics of different gland tissues, we identified 511 gland-specific genes. Of these, 77 genes were functionally examined for gland morphological changes and alterations in gland volatile composition. We showed that 67 genes (87%) cause alterations of at least one gland volatile after RNA interference-based gene knock-down. Three novel genes with quinone-less phenotypes were characterized further to confirm their importance in defensive secretions and their independent evolutionary origin.
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影响因子：
4.1
作者：
Alexander, P;Barton, DHR
通讯作者：
Barton, DHR
DOI：
10.1007/bf01969615
发表时间：
1980-01-01
期刊：
EXPERIENTIA
影响因子：
--
作者：
FAYE, I;WYATT, GR
通讯作者：
WYATT, GR
影响因子：
5.8
作者：
Conesa, A;Götz, S;Robles, M
通讯作者：
Robles, M
DOI：
10.1016/0020-1790(77)90057-9
发表时间：
1977-01-01
期刊：
INSECT BIOCHEMISTRY
影响因子：
--
作者：
DUFFEY, SS;BLUM, MS
通讯作者：
BLUM, MS
影响因子：
1.5
作者：
EISNER, T;MCHENRY, F;SALPETER, MM
通讯作者：
SALPETER, MM