Analysis of gene expression in emerald ash borer (Agrilus planipennis) using quantitative real time-PCR.
Emerald ash borer (EAB, Agrilus planipennis) is an exotic invasive pest, which has killed millions of ash trees (Fraxinus spp) in North America. EAB continues to spread rapidly and attacks ash trees of different ages, from saplings to mature trees. However, to date very little or no molecular knowledge exists for EAB. We are interested in deciphering the molecular-based physiological processes at the tissue level that aid EAB in successful colonization of ash trees. In this report we show the effective use of quantitative real-time PCR (qRT-PCR) to ascertain mRNA levels in different larval tissues (including midgut, fat bodies and cuticle) and different developmental stages (including 1(st)-, 2(nd)-, 3(rd)-, 4(th)-instars, prepupae and adults) of EAB. As an example, a peritrophin gene (herein named, AP-PERI1) is exemplified as the gene of interest and a ribosomal protein (AP-RP1) as the internal control. Peritrophins are important components of the peritrophic membrane/matrix (PM), which is the lining of the insect gut. The PM has diverse functions including digestion and mechanical protection to the midgut epithelium.
Bhandary
Binny
B
Department of Entomology, The Ohio State University.
Rajarapu
Swapna Priya
SP
Rivera-Vega
Loren
L
Mittapalli
Omprakash
O
D000818:Animals; D001517:Coleoptera / Q000235:genetics*; D031661:Fraxinus / Q000469:parasitology; D020869:Gene Expression Profiling / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D012333:RNA, Messenger
Nucleic Acids Res. 2001 May 1;29(9):e45
11328886
Annu Rev Entomol. 1997;42:525-50
15012322
Arch Insect Biochem Physiol. 2007 Jan;64(1):19-29
17167751
Insect Biochem Mol Biol. 2009 May-Jun;39(5-6):403-13
19364528