Jatropha curcas is one of the bio-energy plants with more development potential. In recent years, research on the gene function of Jatropha curcas has become more and more in-depth, but so far, no suitable internal reference genes have been screened for the gene expression analysis of Jatropha curcas. With the wide application of RT-qPCR experimental technology, it is particularly important to select the appropriate internal reference gene as a reference standard when analyzing the expression of genes.
Under the guidance of the tutor Xu Zengfu, Dr. Zhang Lu, a Ph.D. candidate in the Energy Plant Molecular Breeding Group of the Xishuangbanna Tropical Botanical Garden of the Chinese Academy of Sciences, used RT-qPCR technology combined with qBasePLUS analysis software to screen out the suitable candidates for Tungzi from 11 commonly used internal reference genes Combination of internal reference genes in different experimental sample sets (20 samples in total). The results showed that in the concentration of tissue samples at different developmental stages (vegetative growth and reproductive growth), Actin, GAPDH and EF1α were the best combination of internal reference genes; in the concentration of tissue samples under abiotic stress (dehydration stress and cold stress), Actin, GAPDH and TUB5 are the best combination of internal reference genes; in the tissue sample concentration of individual vegetative growth stages, Actin and TUB8 are the best combination of internal reference genes; in the tissue samples of separate reproductive growth stages, GAPDH and EF1α are the best Internal reference gene combination; in the sample set of dehydration stress alone, TUB5 and TUB8 are the best internal reference gene combination; in the sample set of cold stress alone, GAPDH and Actin are the best internal reference gene combination. In addition, the screening results of the internal reference gene combinations in the dehydration stress sample set were also standardized and verified in this study. Different experimental sample sets should select corresponding internal reference gene combinations for gene expression analysis. The results of this study have great reference value for gene expression analysis in the study of Jatropha curcas gene function.
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