Validation of reference genes in leaf-cutting ant Atta sexdens rubropilosa in different developmental stages and tissues

Validation of reference genes in leaf-cutting ant Atta sexdens rubropilosa in different developmental stages and tissues



Ariele C. Moreira, Adriana M. dos Santos, Renato Lajarim Carneiro, Odair C. Bueno, Dulce Helena F. Souza



Atta sexdens rubropilosa is an important leaf-cutting ant species
considered as a pest in agricultural crop or reforestation areas.
Quantitative real-time reverse transcriptase-polymerase chain reaction
(RT-qPCR) is a technique that can help us to understand the regulation
and the function of a gene. However, its reliability depend on the data
normalization. Different normalization strategies can be adopted for
qPCR, reference genes has been cited as one of the most effective
methods. It has not been identified a universal reference for all
organism and experiment. In this way, the validation of reference gene
is crucial step. This is the first study to evaluate reference genes for
leaf-cutting ants. To this, we analyzed the expression levels of
candidate reference genes (act, ef1-alpha, ef1-beta, GAPDH and rpl18) in
different developmental stages (larva, pupa and worker) and tissues
(head, mesosoma and worker without gaster) of A. sexdens rubropilosa.
Four different algorithms (BestKeeper, geNorm, NormFinder and
comparative ΔCt method) were used in statistical analysis of the
stability of the genes and RefFinder was used to propose a consensus
list for ranking the reference genes. Our results showed that the most
suitable combinations of reference gene candidates were rpl18 and
ef1-alpha for the different developmental stages and rpl18 and ef1-beta
for the different tissues. In this work, we also report the obtaining
from a putative acetylcholinesterase from A.sexdens rubropilosa (GenBank
KY464935), which was used as a target gene to confirm the reliability
of reference genes suggested. 



10.22161/ijeab/2.2.23



http://ijeab.com/upload_document/issue_files/23%20IJEAB-MAR-2017-17-Validation%20of%20reference%20genes%20in%20leaf-cutting%20ant.pdf



http://ijeab.com/submit-paper/

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