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Bulletin of Botanical Research ›› 2016, Vol. 36 ›› Issue (4): 565-572.doi: 10.7525/j.issn.1673-5102.2016.04.012

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Selection of Reference Genes for Real-time Fluorescence Quantitative PCR in Different Tissues and Stigma Development from Ornamental Kale

LI Han, LI Zhi-Long, LI Xiao-Yu, LI Yu-Hua, LAN Xing-Guo   

  1. College of Life Sciences, Northeast Forestry University, Harbin 150040
  • Received:2015-12-17 Online:2016-07-15 Published:2016-06-15
  • Supported by:
    Central Universities Fundamental Research Special Fund Project(DL13CA13);National Natural Science Foundation of China(31070275)

Abstract: We used real-time quantitative polymerase chain reaction to test the expression of Actin, cysteine proteinase inhibitor 6(Cpi6), elongation factor 1-beta(EF-1β), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), tubulin alpha-3(Tub-α3), tubulin alpha-6(Tub-α6) and ubiquitin conjugating enzyme7(Ubc7), and then identified the most suitable reference genes in a given set of tissues and different developmental stigmas of ornamental kale(Brassica oleracea var. acephala) S13-bS13-b homozygotes by using the geNorm and BestKeeper software programs. By a geNorm analysis, Tub-α6 and EF-1β were the most suitable reference genes among the given set of tissues, and Actin and Ubc7 were the most stable genes during stigma development. By a BestKeeper analysis, Ubc7 and Tub-α6 were the most suitable reference genes among the given set of tissues, and EF-1β and Tub-α6 were the most stable genes during stigma development. Tub-α6 and Actin were the most suitable reference genes among the given set of tissues and during stigma development, respectively. Furthermore, the expression of SLG normalized with Tub-α6 or Actin showed that SLG was predominantly expressed in stigma among the given set of tissues and reached its highest level at approaching anthesis during stigma development.

Key words: Brassica oleracea var. acephala, real-time quantitative PCR, reference genes, stigma development, SLG

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