globalchange  > 气候变化与战略
DOI: 10.1016/j.plgene.2019.100217
论文题名:
Validation of suitable reference genes for normalization of quantitative reverse transcriptase- polymerase chain reaction in rice infected by Xanthomonas oryzae pv. oryzae
作者: Sagun C.M.; Grandmottet F.; Suachaowna N.; Sujipuli K.; Ratanasut K.
刊名: Plant Gene
ISSN: 23524073
出版年: 2020
卷: 21
语种: 英语
英文关键词: Gene expression ; Housekeeping gene ; Normalization ; qRT-PCR ; Reference gene ; Rice ; Xanthomonas
英文摘要: Bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a costly disease in rice that threatens global rice production. Gene expression analysis by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) allows the study of the underlying mechanisms of both BB pathogenesis and resistance. In relative quantification, reference genes are often used to normalize the results to remove technical variations allowing the determination of true biological changes in a pilot experiment. However, variations in the expression of these reference genes can lead to erroneous and unreliable results. Thus, choosing the most stable reference genes for any specific experimental condition is of utmost importance in qRT-PCR experiments. Here, we used geNorm, NormFinder, Bestkeeper, Delta-Ct and RefFinder programs and/or methods to analyze the stability of the expression of eleven candidate reference genes namely: 18S ribosomal RNA (18S rRNA), Actin-1 (ACT1), ADP-Ribosylation Factor (ARF), Endothelial differentiation factor (Edf), eukaryotic Elongation Factor-1α (eEF-1α), eukaryotic Initiation Factor-4a (eIF-4a), Profilin 2 (Prof2), Nucleic Acid Binding Protein (NABP), Triosephosphate Isomerase (TI), Ubiquitin 5 (UBQ5) and Ubiquitin 10 (UBQ10) in cDNA samples from BB-susceptible and Xa21-mediated resistant rice cultivars collected at various times after Xoo inoculation. Under our experimental conditions, Edf and TI were the most stable reference genes while the common housekeeping genes 18S rRNA, and UBQ5 were among the least stable genes. Though using either Edf or TI as internal control is adequate for gene expression analysis, we suggest using both genes to normalize the data of qRT-PCR assays for rice subjected to Xoo inoculation. © 2019 Elsevier B.V.
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资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/159157
Appears in Collections:气候变化与战略

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作者单位: Department of Agricultural Science, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, Phitsanulok, 65000, Thailand; Center of Excellence in Research for Agricultural Biotechnology, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, Phitsanulok, 65000, Thailand; Department of Research and Development, Tarlac Agricultural University, Malacampa, Camiling, Tarlac 2306, Philippines

Recommended Citation:
Sagun C.M.,Grandmottet F.,Suachaowna N.,et al. Validation of suitable reference genes for normalization of quantitative reverse transcriptase- polymerase chain reaction in rice infected by Xanthomonas oryzae pv. oryzae[J]. Plant Gene,2020-01-01,21
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