Background: RT-qPCR is a preferred method for rapid and reliable quantification of gene expression studies. Appropriate application of RT-qPCR in such studies requires the use of reference gene(s) as an internal control to normalize mRNA levels between different samples for an exact comparison of gene expression level. However, recent studies have shown that no single reference gene is universal for all experiments. Thus, the identification of high quality reference gene(s) is of paramount importance for the interpretation of data generated by RT-qPCR. Only a few studies on reference genes have been done in plants and none in peach (Prunus persica L. Batsch). Therefore, the present study was conducted to identify suitable reference gene(s) for normalization of gene expression in peach. Results: In this work, eleven reference genes were investigated in different peach samples using RT-qPCR with SYBR green. These genes are: actin 2/7 (ACT), cyclophilin (CYP2), RNA polymerase II (RP II), phospholipase A2 (PLA2), ribosomal protein L13 (RPL13), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S ribosomal RNA (18S rRNA), tubblin beta (TUB), tubblin alpha (TUA), translation elongation factor 2 (TEF2) and ubiquitin 10 (UBQ10). All eleven reference genes displayed a wide range of C_q values in all samples, indicating that they expressed variably. The stability of these genes except for RPL13 was determined by three different descriptive statistics, geNorm, NormFinder and BestKeeper, which produced highly comparable results. Conclusion: Our study demonstrates that expression stability varied greatly between genes studied in peach. Based on the results from geNorm, NormFinder and BestKeeper analyses, for all the sample pools analyzed, TEF2, UBQ10 and RP II were found to be the most suitable reference genes with a very high statistical reliability, and TEF2 and RP II for the other sample series, while 18S rRNA, RPL13 and PLA2 were unsuitable as internal controls. GAPDH and ACT also performed poorly and were less stable in our analysis. To achieve accurate comparison of levels of gene expression, two or more reference genes must be used for data normalization. The combinations of TEF2/UBQ10/RP II and TEF2/RP II were suggested for use in all samples and subsets, respectively.

Hormones are closely associated with dormancy in deciduous fruit trees, and gibberellins (GAs) are known to be particularly important. In this study, we observed that GA₄ treatment led to earlier bud break in Japanese apricot. To understand better the promoting effect of GA₄ on the dormancy release of Japanese apricot flower buds, proteomic and transcriptomic approaches were used to analyse the mechanisms of dormancy release following GA₄ treatment, based on two-dimensional gel electrophoresis (2-DE) and digital gene expression (DGE) profiling, respectively. More than 600 highly reproducible protein spots (P<0.05) were detected and, following GA₄ treatment, 38 protein spots showed more than a 2-fold difference in expression, and 32 protein spots were confidently identified according to the databases. Compared with water treatment, many proteins that were associated with energy metabolism and oxidation–reduction showed significant changes after GA₄ treatment, which might promote dormancy release. We observed that genes at the mRNA level associated with energy metabolism and oxidation–reduction also played an important role in this process. Analysis of the functions of the identified proteins and genes and the related metabolic pathways would provide a comprehensive proteomic and transcriptomic view of the coordination of dormancy release after GA₄ treatment in Japanese apricot flower buds.

Gibberellin (GA₄) has a significant effect on promoting dormancy release in flower buds of Japanese apricot (Prunus mume Sieb. et Zucc). The transcriptomic and proteomic changes that occur after GA₄ treatment have been reported previously; however, the metabolic changes brought about by GA₄ remain unknown. The present study was undertaken to assess changes in metabolites in response to GA₄ treatment, as determined using gas chromatography–mass spectrometry and principal component analysis. Fifty-five metabolites that exhibited more than two-fold differences in abundance (P < 0.05) between samples collected over time after a given treatment or between samples exposed to different treatments were studied further. These metabolites were categorized into six main groups: amino acids and their isoforms (10), amino acid derivatives (7), sugars and polyols (14), organic acids (12), fatty acids (4), and others (8). All of these groups are involved in various metabolic pathways, in particular galactose metabolism, glyoxylate and dicarboxylate metabolism, and starch and sucrose metabolism. These results suggested that energy metabolism is important at the metabolic level in dormancy release following GA₄ treatment. We also found that more than 10-fold differences in abundance were observed for many metabolites, including sucrose, proline, linoleic acid, and linolenic acid, which might play important roles during the dormancy process. The current research extends our understanding of the mechanisms involved in budburst and dormancy release in response to GA₄ and provides a theoretical basis for applying GA₄ to release dormancy.

MicroRNAs (miRNAs) are a class of endogenous, small, non-coding RNAs that regulate gene expression by mediating gene silencing at transcriptional and post-transcriptional levels in high plants. However, the diversity of miRNAs and their roles in floral development in Japanese apricot (Prunus mume Sieb. et Zucc) remains largely unexplored. Imperfect flowers with pistil abortion seriously decrease production yields. To understand the role of miRNAs in pistil development, pistil development-related miRNAs were identified by Solexa sequencing in Japanese apricot.

The strawberry (Fragaria × ananassa Duch.) is an important fruit crop cultivated worldwide for its unique taste and nutritional properties. One of the major risks associated with strawberry production is cold damage. Recently, melatonin has emerged as a multifunctional signaling molecule that influences plant growth and development and reduces adverse consequences of cold stress. The present study was conducted to investigate the defensive role of melatonin and its potential interrelation with abscisic acid (ABA) in strawberry plants under cold stress. The results demonstrate that melatonin application conferred improved cold tolerance on strawberry seedlings by reducing malondialdehyde and hydrogen peroxide contents under cold stress. Conversely, pretreatment of strawberry plants with 100 μM melatonin increased soluble sugar contents and different antioxidant enzyme activities (ascorbate peroxidase, catalase, and peroxidase) and non-enzymatic antioxidant (ascorbate and glutathione) activities under cold stress. Furthermore, exogenous melatonin treatment stimulated the expression of the DREB/CBF—COR pathways’ downstream genes. Interestingly, ABA treatment did not change the expression of the DREB/CBF—COR pathway. These findings imply that the DREB/CBF-COR pathway confers cold tolerance on strawberry seedlings through exogenous melatonin application. Taken together, our results reveal that melatonin (100 μM) pretreatment protects strawberry plants from the damages induced by cold stress through enhanced antioxidant defense potential and modulating the DREB/CBF—COR pathway.