Two microRNA (miRNA) quantification methods, namely, poly(A) reverse transcription (RT)-quantitative real-time polymerase chain reaction (qPCR) and stem-loop RT-qPCR, have been developed for quantifying miRNA expression. In the present study, five miRNAs, including miR166, miR167, miR168, miR159, and miR396, with different sequence frequencies, were selected as targets to compare their expression profiles in five wheat tissues by applying the two methods and deep sequencing. The study aimed to determine a simple, reliable and high-throughput method for detecting miRNA expressions in wheat tissues. Results showed that the miRNA expression profiles determined by poly(A) RT-qPCR were more consistent with those obtained by deep sequencing. Further analysis indicated that the correlation coefficients of the data obtained by poly(A) RT-qPCR and deep sequencing (0.739, P 0.01) were higher than those obtained by stem-loop RT-qPCR and deep sequencing (0.535, P 0.01). The protocol used for poly(A) RT-qPCR is simpler than that for stem-loop RT-qPCR. Thus, poly(A) RT-qPCR was a more suitable high-throughput assay for detecting miRNA expression profiles. To the best of our knowledge, this study was the first to compare these two miRNA quantification methods. We also provided useful information for quantifying miRNA in wheat or other plant species.

The soil organic matter and nutrients are fundamental for the sustainability of pear production, but little is known about the spatial distribution of soil organic matter and nutrients in a pear orchard. With the soil of the pear (cv. Dangshansu on P.betulifolia Bunge. rootstock) orchard under clean and sod cultivation models as test materials, the experiment was conducted to evaluate spatial variability of soil organic matter (SOM), total nitrogen (STN), total phosphorus (STP), total potassium (STK), available nitrogen (SAN), and available potassium (SAK) in and between rows at different soil depths (0-60 cm). The SOM, STN, STP, STK, SAN and SAK of the different soil layers under the two tillage models were different in the vertical direction. The SOM, STN, STP and SAN in the 0-20 cm soil layer were higher than those in the 20-40 and 40- 60 cm soil layers. The STK of 40-60 cm soil layer was higher than that in the 0-20 and 20-40 cm soil layers. The STK increased with the depth of soil in the vertical direction in the clean cultivated pear orchard. Variability of the SOM, STN, STP, STK, SAN and SAK of sample sites in between rows of the same soil layer was found in the pear orchard soil in the horizontal direction under clean and sod cultivation management systems, except that STK of all sites did not show the difference in identical soil layers in the pear orchard under clean cultivation. The sod cultivation model improved the SOM, STN, and STK in the 0-20 cm soil layer in the pear orchard, and the three components increased by 12.8, 12.7 and 7.3% compared to clean cultivation, respectively. The results can be applicable to plan collection of orchard soil samples, assess orchard soil quality, and improve orchard soil management practices.