Li, Q. and Fan, C-M. and Zhang, X-M. and Fu, Y-F. (2012) Validation of reference genes for real-time quantitative PCR normalization in soybean developmental and germinating seeds. Plant Cell Reports. 10 p..
![[img]](http://eprints.icrisat.ac.in/style/images/fileicons/application_pdf.png) |
PDF (Online First)
- Published Version
Restricted to ICRISAT researchers only |
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 Glyma05g 37470, Glyma08g28550, Glyma18g04130 and CYP for normalization.
| Item Type: | Article |
|---|---|
| Additional Information: | This work was partly supported by Transgenic program (Nos 2009ZX08009-133B, 2011ZX08009-001 and 2011ZX08004-005), the Chinese National Key Basic Research ‘‘973’’ Program (2010CB125906), and the National Natural Science Founds (31000680 and 31000681). |
| Uncontrolled Keywords: | Soybean, Seed development, Seed germination, RT-qPCR, Normalization, Reference gene |
| Author Affiliation: | 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 |
| Subjects: | Crop Improvement |
| Divisions: | Soyabean |
| Depositing User: | Mr Siva Shankar |
| Date Deposited: | 08 Aug 2012 03:52 |
| Last Modified: | 08 Aug 2012 03:53 |
| Official URL: | http://dx.doi.org/10.1007/s00299-012-1282-4 |
| URI: | http://eprints.icrisat.ac.in/id/eprint/7227 |
Actions (login required)
 |
View Item |
