PMID- 31877150
OWN - NLM
STAT- MEDLINE
DCOM- 20200327
LR  - 20200327
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 14
IP  - 12
DP  - 2019
TI  - Selection of valid reference genes for quantitative real-time PCR in Cotesia 
      chilonis (Hymenoptera: Braconidae) exposed to different temperatures.
PG  - e0226139
LID - 10.1371/journal.pone.0226139 [doi]
LID - e0226139
AB  - In quantitative real-time PCR (qRT-PCR), data are normalized using reference 
      genes, which helps to control for internal differences and reduce error among 
      samples. In this study, the expression profiles of eight candidate housekeeping 
      genes, 18S ribosomal (18S rRNA), elongation factor (EF1), 
      glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein L10 (RPL10), 
      ribosomal protein L17 (RPL17), histone 3 (H3), arginine kinase (AK), amd beta-Actin 
      (ACTB), were evaluated in the parasitic wasp Cotesia chilonis in response to 
      different temperatures. Specifically, the performance and stabilities of these 
      genes were compared in adult wasps maintained in a growth condition at 27 degrees C 
      (normal storage conditions) and in adults obtained from pupae refrigerated at 4 degrees C 
      for five days (cold storage conditions). Data were analyzed using the DeltaCt method, 
      BestKeeper, NormFinder, and geNorm. The optimal numbers and stabilities of 
      reference genes varied between the two temperature treatments (27 degrees C and 4 degrees C). In 
      samples stored at normal developmental temperature (27 degrees C), the requirement for 
      normalization in response to low temperature exposures was three genes (18S, H3, 
      AK), whereas normalization in response to high temperature exposures required 
      only two reference genes (GAPDH, ACTB). In samples stored at cold temperature 
      (4 degrees C), for low temperature exposures two reference genes (RPL17, RPL10) were 
      required for standardization, while following high temperature exposures three 
      reference genes (18S, H3, ACTB) were needed. This study strengthens understanding 
      of the selection of reference genes before qRT-PCR analysis in C. chilonis. The 
      reference genes identified here will facilitate further investigations of the 
      biological characteristics of this important parasitoid.
FAU - Li, Qiu-Yu
AU  - Li QY
AD  - School of Horticulture and Plant Protection & Institute of Applied Entomology, 
      Yangzhou University, Yangzhou, China.
FAU - Li, Zi-Lan
AU  - Li ZL
AD  - School of Horticulture and Plant Protection & Institute of Applied Entomology, 
      Yangzhou University, Yangzhou, China.
FAU - Lu, Ming-Xing
AU  - Lu MX
AD  - School of Horticulture and Plant Protection & Institute of Applied Entomology, 
      Yangzhou University, Yangzhou, China.
AD  - Joint International Research Laboratory of Agriculture and Agri-Product Safety, 
      the Ministry, Yangzhou, China.
FAU - Cao, Shuang-Shuang
AU  - Cao SS
AD  - School of Horticulture and Plant Protection & Institute of Applied Entomology, 
      Yangzhou University, Yangzhou, China.
FAU - Du, Yu-Zhou
AU  - Du YZ
AUID- ORCID: 0000-0002-4452-7125
AD  - School of Horticulture and Plant Protection & Institute of Applied Entomology, 
      Yangzhou University, Yangzhou, China.
AD  - Joint International Research Laboratory of Agriculture and Agri-Product Safety, 
      the Ministry, Yangzhou, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191226
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Actins)
RN  - 0 (Histones)
RN  - 0 (Insect Proteins)
RN  - 0 (Peptide Elongation Factor 1)
RN  - 0 (RNA, Ribosomal, 18S)
RN  - 0 (RPL10 protein, human)
RN  - 0 (Ribosomal Proteins)
RN  - 0 (ribosomal protein L17)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
RN  - EC 2.7.3.3 (Arginine Kinase)
SB  - IM
MH  - Actins/genetics
MH  - Animals
MH  - Arginine Kinase/genetics
MH  - Gene Expression Profiling/*standards
MH  - *Genes, Essential
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/genetics
MH  - Histones/genetics
MH  - Hot Temperature
MH  - Insect Proteins/genetics
MH  - Peptide Elongation Factor 1/genetics
MH  - RNA, Ribosomal, 18S/genetics
MH  - Real-Time Polymerase Chain Reaction/*standards
MH  - Reference Standards
MH  - Ribosomal Protein L10/genetics
MH  - Ribosomal Proteins/genetics
MH  - Stress, Physiological
MH  - Wasps/genetics/*physiology
PMC - PMC6932786
COIS- The authors have declared that no competing interests exist.
EDAT- 2019/12/27 06:00
MHDA- 2020/03/28 06:00
PMCR- 2019/12/26
CRDT- 2019/12/27 06:00
PHST- 2019/05/05 00:00 [received]
PHST- 2019/11/20 00:00 [accepted]
PHST- 2019/12/27 06:00 [entrez]
PHST- 2019/12/27 06:00 [pubmed]
PHST- 2020/03/28 06:00 [medline]
PHST- 2019/12/26 00:00 [pmc-release]
AID - PONE-D-19-11577 [pii]
AID - 10.1371/journal.pone.0226139 [doi]
PST - epublish
SO  - PLoS One. 2019 Dec 26;14(12):e0226139. doi: 10.1371/journal.pone.0226139. 
      eCollection 2019.