Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (5): 955-963.DOI: 10.3969/j.issn.1000-2561.2020.05.015

• Biotechnology and Tissue Culture • Previous Articles     Next Articles

Screening of Reference Genes for Quantitative Real-time PCR of Rubber Saplings under Aluminum Stress

MA Xiaowei1,2,AN Feng2,*(),LIU Zifan1,*(),XIE Guishui2   

  1. 1. College of Tropical Crops, Hainan University, Haikou, Hainan 570228, China
    2. Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences / Danzhou Investigation & Experiment Station of Tropical Crops, Ministry of Agriculture & Rural Affairs, Danzhou, Hainan 571737, China
  • Received:2019-08-15 Revised:2019-09-08 Online:2020-05-25 Published:2020-06-15
  • Contact: AN Feng,LIU Zifan


Aluminum toxicity is a major factor inhibiting crop growth on tropical acidic soils. However, the study on the effects of aluminum toxicity on rubber tree growth and latex production is limited, the mechanism of rubber trees tolerate to aluminum has not been reported. To screen the reference genes stably expressed in rubber trees while aluminum stress and elucidate the molecular mechanism of rubber tree aluminum tolerance by real-time quantitative PCR assay, ten candidate internal reference genes, i.e. Actin, Actin7, 18S rRNA, 40S rRNA, YLS8, UBC2, UBC4, GAPDH, FP and ADF were selected in this study. The expression stability was analyzed with geNorm, NormFinder, BestKeeper and Delta Ct and RefFinder. The expression stability of the ten candidate internal reference genes were different. The top three stable reference genes were UBC4, 40S rRNA and FP. Meanwhile, the expression of two aluminum-induced malate transporter genes (HbALMT-1 and HbALMT-2) in rubber trees were verified with UBC4, 40S rRNA, FP, the combination of top three stable genes (UBC4+40S rRNA+FP) and the unstable gene GAPDH. The expression profiles of the two target genes were consistent when normalized by three reference genes and the combinations. The unstable reference gene (GAPDH) failed to standardize the expression data. In conclusion, UBC4, 40S rRNA and FP genes and the combinations were selected as the most stable reference genes, which could be normalized the expression of the relative genes under aluminum stress.

Key words: rubber tree, aluminum toxicity, quantitative real-time PCR, reference gene

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