Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2013, Vol. 29 ›› Issue (10): 11-17.doi: 10.11924/j.issn.1000-6850.2012-3872

Special Issue: 生物技术

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Reference Genes Selection for Real-time fluorescence quantitative PCR in L. chinense var. rubrum

  

  • Received:2012-11-29 Revised:2013-01-06 Online:2013-04-05 Published:2013-04-05

Abstract: Real-time fluorescence quantitative PCR(qRT-PCR) has been widely used in gene expression analysis to date, and selection of suitable reference genes for qRT-PCR according to specific experimental materials or conditions is very important for accurate normalization of target gene expression. However, the studies on reference genes have not been done in Loropetalurn, In this study, expression of four commonly used housekeeping genes such as α-tubulin , β-actin ,18S ribsomal RNA (18S rRNA) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were assessed by qRT-PCR in various tissues (buds, young stems, leaves, petals ,seeds and callus) . All the candidate reference genes could amplify specifically with high efficiency in qRT-PCR reaction. To determine the expression stability of these genes, the data were determined by two commonly used applets(geNorm and NormFinder), which produced highly comparable results depending on the experimental parameters. GeNorm and NormFinder algorithms revealed that β-actin and GAPDH were both in the highest stability, and GAPDH could be used as a reference gene for various organs and tissues and β-actin for leaves at different stages and callus among diverse treatments.

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