IDENTIFICATION OF MYCOSIS-RELATED GENES IN THE EASTERN SUBTERRANEAN TERMITE BY SUPPRESSION SUBTRACTIVE HYBRIDIZATION
IDENTIFICATION OF MYCOSIS-RELATED GENES IN THE EASTERN SUBTERRANEAN TERMITE BY SUPPRESSION SUBTRACTIVE HYBRIDIZATION
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DOI：
10.1002/arch.21026
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发表时间：
2012-07-01
影响因子：
2.2
通讯作者：
Thompson, Graham J.
中科院分区：
文献类型：
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作者：
Gao, Qi;Tancredi, Sarah E.;Thompson, Graham J.
东部地下白蚁（等翅目，鼻白蚁科）是一种世界性的结构性害虫，是白蚁先天免疫研究的目标。在这项研究中，我们使用抑制减法杂交构建了一个标准化的cDNA文库，该文库包含真菌感染时过度表达的基因。在感染绿僵菌24小时后，文库发现182个表达序列标签（EST）克隆可能代表免疫应答基因。大多数（97%）ESTs的核苷酸序列组装成少数（n = 13）个连续序列，其余（n = 6）为单核苷酸序列。因此，我们的筛选捕获了多达19种不同的mrna，这些mrna在对真菌病原体的反应中高度表达。所有基因座的初步测序显示，大约一半（n = 10）包含与已知蛋白显著相似的开放阅读框。这些克隆代表核和线粒体编码基因，以及假定的长链非编码RNA基因。对独立感染工蚁群体的编码基因进行定量聚合酶链反应分析，证实在每种情况下，从文库中鉴定的转录本在真菌感染后都是上调的。这些基因对今后白蚁生物防治和群居昆虫免疫的研究具有重要意义。
The Eastern subterranean termite Reticulitermes flavipes (Isoptera, Rhinotermitidae) is a cosmopolitan, structural pest that is the target of research into termite innate immunity. In this study, we use suppression subtractive hybridization to construct a normalized cDNA library of genes excessively expressed upon fungal infection. At 24 h postinfection with Metarhizium anisopliae, the library revealed 182 expressed sequence tag (EST) clones that potentially represent immune responsive genes. The nucleotide sequence from a majority (97%) of ESTs assembled into a small number (n = 13) of contiguous sequences, with the remainder (n = 6) representing singletons. Our screen therefore captured as many as 19 different mRNAs highly expressed in response to the fungal pathogen at this time. Primary sequencing of all loci revealed that approximately half (n = 10) contained open reading frames with significant similarity to known proteins. These clones represent nuclear and mitochondrial coding genes, as well as putative long noncoding RNA genes. Quantitative polymerase chain reaction analysis of coding genes on independently infected groups of worker termites confirms in each case that the transcripts identified from the library are up-regulated postfungal infection. The genes identified here are relevant to future studies on termite biocontrol and social insect immunity.