There are no comprehensive methods to identify N6-methyladenosine (m6A) at single-base resolution for every single transcript, which is necessary for the estimation of m6A abundance. We develop a new pipeline called Nanom6A for the identification and quantification of m6A modification at single-base resolution using Nanopore direct RNA sequencing based on an XGBoost model. We validate our method using methylated RNA immunoprecipitation sequencing (MeRIP-Seq) and m6A-sensitive RNA-endoribonuclease–facilitated sequencing (m6A-REF-seq), confirming high accuracy. Using this method, we provide a transcriptome-wide quantification of m6A modification in stem-differentiating xylem and reveal that different alternative polyadenylation (APA) usage shows a different ratio of m6A.
High-yield rice cultivation is an effective way to address the increasing food demand worldwide. Correct classification of high-yield rice is a key step of breeding. However, manual measurements within breeding programs are time consuming and have high cost and low throughput, which limit the application in large-scale field phenotyping. In this study, we developed an accurate large-scale approach and presented the potential usage of hyperspectral data for rice yield measurement using the XGBoost algorithm to speed up the rice breeding process for many breeders. In total, 13 japonica rice lines in regional trials in northern China were divided into different categories according to the manual measurement of yield. Using an Unmanned Aerial Vehicle (UAV) platform equipped with a hyperspectral camera to capture images over multiple time series, a rice yield classification model based on the XGBoost algorithm was proposed. Four comparison experiments were carried out through the intraline test and the interline test considering lodging characteristics at the midmature stage or not. The result revealed that the degree of lodging in the midmature stage was an important feature affecting the classification accuracy of rice. Thus, we developed a low-cost, high-throughput phenotyping and nondestructive method by combining UAV-based hyperspectral measurements and machine learning for estimation of rice yield to improve rice breeding efficiency.
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