Background
Before studying gene expression of different organisms, it is important to determine the best reference gene. At present, the most accurate method of detecting gene expression is quantitative real-time PCR (RT-qPCR). With this method, reference genes that are stable in different biological systems and under different conditions can be obtained. Toona ciliata Roem (T. ciliata). is a valuable and fast-growing timber specie. In this study, 20 reference genes were identified using RT-qPCR, as a primary prerequisite for future gene expression analysis. Four different methods, geNorm, NormFinder, BestKeeper, and RankAggreg were used to evaluate the expression stability of the 20 candidate reference genes in various tissues under different conditions.
Results
The experimental results showed that TUB-α was the most stably expressed reference gene across all samples and UBC17 was the most stable in leaves and young stems under Hypsipyla robusta (H. robusta) and methyl jasmonate (MeJA) treatments. In addition, PP2C59 and UBC5B were the best-performing genes in leaves under H. robusta treatment, while HIS1 and ACT7 were the best reference genes in young stems. The two best reference genes were 60S-18 and TUB-α after treatment at 4 °C. The expression of HIS6 and MUB1 was the most stable under PEG6000 treatment. The accuracy of the selected reference genes was verified using the transcription factor MYB3 (TcMYB3) gene.
Conclusions
This is the first report to verify the best reference genes for normalizing gene expression in T. ciliata under different conditions, which will facilitate future elucidation of gene regulations in this species.
Analyzing the temporal variation of runoff and vegetation and quantifying the impact of anthropic factors and climate change on vegetation and runoff variation in the source area of the Yangtze River (SAYR), is of great significance for the scientific response to the ecological protection of the region. Therefore, the Budyko hypothesis method and multiple linear regression method were used to quantitatively calculate the contribution rates of climate change and anthropic factors to runoff and vegetation change in the SAYR. It was found that: (1) The runoff, NDVI, precipitation, and potential evaporation in the SAYR from 1982 to 2016 all showed an increasing trend. (2) The mutation year of runoff data from 1982 to 2016 in the SAYR is 2004, and the mutation year of NDVI data from 1982 to 2016 in the SAYR is 1998. (3) The contribution rates of precipitation, potential evaporation and anthropic factors to runoff change of the SAYR are 75.98%, −9.35%, and 33.37%, respectively. (4) The contribution rates of climatic factors and anthropic factors to vegetation change of the SAYR are 38.56% and 61.44%, respectively.
Neolamarckia cadamba (Roxb.) Bosser is a tropical evergreen broadleaf tree species that could play an important role in meeting the increasing demand for wood products. However, multi-level genetic variation and selection efficiency for growth traits in N. cadamba is poorly characterized. We therefore investigated the efficiency of early selection in N. cadamba by monitoring the height (HT), diameter at breast height (DBH), and tree volume (V) in 39 half-sib families from 11 provenances at ages 2, 3, 4, 5, and 6 years in a progeny test. Age-related trends in growth rate, genetic parameters in multi-level, efficiency of early selection, and realized gain in multi-level for growth traits were analyzed. The result showed that genetic variation among families within provenances was higher than that among provenances. The estimated individual heritability values for the growth traits ranged from 0.05 to 0.26, indicating that the variation of growth traits in N. cadamba was subject to weak or intermediate genetic control. The age–age genetic correlations for growth traits were always positive and high (0.51–0.99), and the relationships between the genetic/phenotypic correlations and the logarithm of the age ratio (LAR) were described well by linear models (R2 > 0.85, except the fitting coefficient of genetic correlation and LAR for HT was 0.35). On the basis of an early selection efficiency analysis, we found that it is the best time to perform early selection for N. cadamba at age 5 before half-rotation, and the selection efficiencies were 157.28%, 151.56%, and 127.08% for V, DBH, and HT, respectively. Higher realized gain can be obtained by selecting superior trees from superior families. These results can be expected to provide theoretical guidance and materials for breeding programs in N. cadamba and can even be a reference for breeding strategies of other fast-growing tree species.
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