Validation of endogenous reference genes for insecticide-induced and developmental expression profiling of Liposcelis bostsrychophila (Psocoptera: Liposcelididae).
Four housekeeping genes named Lbbeta-Actin1, Lbbeta-Actin2, Lbalpha-Tubulin, and LbGapdh were cloned from Liposcelis bostrychophila using the combined techniques of reverse transcriptase-PCR (RT-PCR) with rapid amplification of cDNA ends (RACE). The GenBank accession numbers were FJ196622, FJ447483, FJ595242, and FJ595241, respectively. The full-length cDNA of Lbbeta-Actin1 was a 1,772 bp sequence with an open reading frame (ORF) encoded 376 amino acids, while Lbbeta-Actin2 was 1,350 bp in length containing an ORF encoded 376 amino acids. Furthermore, the 1,565 bp cDNA of Lbalpha-Tubulin had an ORF of 1,350 bp encoding 450 amino acids. And LbGapdh possessed an ORF of 333 amino acids. Sequences analysis and phylogenetic trees generated from the nucleotide sequences of their coding regions revealed a relationship that was closer to other insects than to mammals. The four genes together with 18S rRNA were quantified for transcription stability in L. bostrychophila, and the geNorm software ranked from the most to least were Lbbeta-Actin1 > LbGapdh > Lbalpha-Tubulin > Lbbeta-Actin2 > Lb18S rRNA for deltamethrin induction, while ranked Lbbeta-Actin1 > Lbalpha-Tubulin > Lbbeta-Actin2 > LbGapdh > Lb18S rRNA for the different developmental stages.
Jiang
Hong-Bo
HB
Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, 400716 Chongqing, People's Republic of China.
Liu
Yong-Hua
YH
Tang
Pei-An
PA
Zhou
An-Wei
AW
Wang
Jin-Jun
JJ
D000818:Animals; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D017344:Genes, Insect / Q000187:drug effects; D007300:Insect Control / Q000379:methods / Q000592:standards; D007313:Insecta / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D007305:Insecticide Resistance / Q000235:genetics; D007306:Insecticides / Q000494:pharmacology*; D008018:Life Cycle Stages / Q000187:drug effects / Q000235:genetics; D009570:Nitriles / Q000494:pharmacology; D010802:Phylogeny; D011722:Pyrethrins / Q000494:pharmacology; D012015:Reference Standards; D017422:Sequence Analysis, DNA; D015854:Up-Regulation / Q000187:drug effects*
D007306:Insecticides; D009570:Nitriles; D011722:Pyrethrins; C017180:decamethrin