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PMID: 22588479 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Validation Study

Validation of reference genes for real-time quantitative PCR normalization in soybean developmental and germinating seeds.

Plant cell reports ·Vol. 31 ·No. 10 ·2012-10-00 ·Pages 1789-98

Li Q, Fan CM, Zhang XM, Fu YF

Abstract

Most of traditional reference genes chosen for real-time quantitative PCR normalization were assumed to be ubiquitously and constitutively expressed in vegetative tissues. However, seeds show distinct transcriptomes compared with the vegetative tissues. Therefore, there is a need for re-validation of reference genes in samples of seed development and germination, especially for soybean seeds. In this study, we aimed at identifying reference genes suitable for the quantification of gene expression level in soybean seeds. In order to identify the best reference genes for soybean seeds, 18 putative reference genes were tested with various methods in different seed samples. We combined the outputs of both geNorm and NormFinder to assess the expression stability of these genes. The reference genes identified as optimums for seed development were TUA5 and UKN2, whereas for seed germination they were novel reference genes Glyma05g37470 and Glyma08g28550. Furthermore, for total seed samples it was necessary to combine four genes of Glyma05g37470, Glyma08g28550, Glyma18g04130 and UKN2 [corrected] for normalization. Key message We identified several reference genes that stably expressed in soybean seed developmental and germinating processes.

MeSH Terms
DNA, Complementary/genetics Gene Expression Regulation, Plant Genes, Plant Genomic Instability Germination Oligonucleotide Array Sequence Analysis/methods RNA, Plant/genetics Real-Time Polymerase Chain Reaction/methods,standards Reference Standards Reverse Transcriptase Polymerase Chain Reaction/methods Seeds/genetics,growth & development Soybeans/genetics,growth & development Time Factors Transcriptome
Chemicals
DNA, Complementary RNA, Plant
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Li Qing
MOA Key Lab of Soybean Biology (Beijing), National Key Facility of Crop Gene Resource and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, People's Republic of China. [email protected]
Fan Cheng-Ming
Zhang Xiao-Mei
Fu Yong-Fu
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Article Info
Journal
Plant cell reports
Abbr.
Plant Cell Rep
ISSN
1432-203X
DOI
10.1007/s00299-012-1282-4
Email
[email protected]
Published
2012-10-00
Epub
2012-00-16
Pages
1789-98
Language
English
Region
Germany
NLM ID
9880970
Subset
IM
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1007/s00299-012-1282-4 View journal details
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