Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (3): 504-512.DOI: 10.3969/j.issn.1000-2561.2020.03.012

• Biotechnology and Tissue Culture • Previous Articles     Next Articles

Selection of Reference Genes for Normalization of Quantitative Real-time PCR Analysis in Secondary Laticifer Differentiation Induced by Trichostatin A in Rubber Tree (Hevea brasiliensis Muell. Arg.)

WU Shaohua,ZHANG Shixin,YANG Shuguang,TIAN Weimin()   

  1. Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture and Rural Affairs / State Key Laboratory Breeding Base of Cultivation and Physiology for Tropical Crops, Haikou, Hainan 571101, China
  • Received:2019-05-06 Revised:2019-08-28 Online:2020-03-25 Published:2020-04-16
  • Contact: TIAN Weimin

Abstract:

The identification of differentially expressed genes associated with laticifer differentiation is crucial for understanding the molecular mechanism of laticifer differentiation. The selection of a suitable reference gene is a precondition for accurate gene expression analysis by quantitative real-time PCR (qRT-PCR). In the present study, the expression stability of 22 candidate reference genes were evaluated on the basis of TSA-induced secondary laticifer differentiation using geNorm and NormFinder algorithms. According to the analysis of geNorm and NormFinder, UBC3, UBC4, eIF1Aa were the top three stable genes and ROC3, PTP, CYP2 were the last three stable genes in the process of TSA-induced secondary laticifer differentiation. The expression patterns of HDA1 and HDA2 in TSA treated bark were analyzed based on UBC3, Actin and ROC3 as the reference gene respectively. The results showed that UBC3 could serve as a qRT-PCR reference gene to analyze the gene expression pattern in TSA-induced secondary laticifer differentiation. And ROC3 was not suitable as reference genes for qRT-PCR.

Key words: Hevea brasiliensis Muell. Arg., laticifer differentiation, trichostatin A, reference genes

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