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IDENTIFICATION OF MYCOSIS‐RELATED GENES IN THE E ASTERN SUBTERRANEAN TERMITE BY SUPPRESSION SUBTRACTIVE HYBRIDIZATION
Author(s) -
Gao Qi,
Tancredi Sarah E.,
Thompson Graham J.
Publication year - 2012
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21026
Subject(s) - biology , suppression subtractive hybridization , gene , genetics , cdna library , expressed sequence tag , metarhizium anisopliae , complementary dna , rhinotermitidae , polymerase chain reaction , metarhizium , botany , conidium
The E astern subterranean termite R eticulitermes flavipes ( I soptera, R hinotermitidae) 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 c DNA library of genes excessively expressed upon fungal infection. At 24 h postinfection with M etarhizium anisopliae, the library revealed 182 expressed sequence tag ( EST ) clones that potentially represent immune responsive genes. The nucleotide sequence from a majority (97%) of EST s 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 m RNA s 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.