Stepwise Optimization of the RT-qPCR Protocol and the Evaluation of Housekeeping Genes in Pears (Pyrus bretschneideri) under Various Hormone Treatments and Stresses

Real-time quantitative PCR (RT-qPCR) has become a widely used method for exploring plant gene expression level. The requires using some stably expressed genes as reference to accurately normalize the RT-qPCR data. However, under various stresses and hormone treatments, levels of most vary. Environmental variations also influence their levels. lack validated, can mislead study function in pears. “Huangguan” pears have recently focus research on stress resistance mechanisms, such high resistance. Therefore, aim this was select optimal Huangguan pears, we analyzed EF1α, ACT, SKD1, YLS8, UBQ, GAPDH, TUB, WDP series pear leaf sets treatments. Using different statistical algorithms, found that had more stable expression, ACT MeJA treatment, YLS8 valuable ABA heat conditions, GAPDH showed worst results compared other housekeeping genes, except stress. These will supply updated information selection biotic abiotic future.