Journal
PLANT SCIENCE
Volume 178, Issue 4, Pages 359-365Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.plantsci.2010.02.005
Keywords
Reference genes; Longan somatic embryogenesis; qPCR; Normalization
Categories
Funding
- National Science and Technology Supporting Project [2007BAD07B01]
- National Natural Science Foundation of China [30471204]
- Fok Ying Tung Education Foundation [71026]
- Higher Education of Chinese Ministry of Education [20093515110006]
- Fujian Provincial Science and Technology Platform Construction Project [2008N2001]
- Fujian Provincial Special Program for Key Science and Technology [2004NZ02-2]
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Real-time reverse transcriptase PCR is a powerful tool to investigate relevant changes in gene expression during plant somatic embryogenesis (SE.): however, this method lacks ideal reference genes. To select the most stable reference genes for S.E. studies, the expression profiles of seven frequently used reference genes (18S RNA, eIF-4a, UBQ ACTB, EF-1a, Histone H3, and 2-TUB) and functional genes (Fe-SOD. Cu/Zn-SOD, and Mn-SOD) were tested in synchronized longan tree embryogenic cultures at different developmental stages at temperatures of 20 degrees C, 25 degrees C, and 30 degrees C. The expression of the 10 candidate genes showed little variation during longan S.E. at 25 degrees C. The most stable combination for gene expression analysis was UBQ and EF-1a or Fe-SOD and UBQ. However, the expression of these genes varied considerably at different developmental stages at different temperatures. Comprehensive analysis of the results using the software packages geNorm, BestKeeper, and NormFinder revealed that UBQ and Fe-SOD together could be used as internal controls for gene expression analysis in a wide variety of stages of longan S.E. cultured under different temperatures. (C) 2010 Elsevier Ireland Ltd. All rights reserved.
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