Plants have evolved complex mechanisms to adapt to harsh environmental conditions. Rice (Oryza sativa) is a staple food crop that is sensitive to low temperatures. However, its cold stress responses remain poorly understood, thus limiting possibilities for crop engineering to achieve greater cold tolerance. In this study, we constructed a rice pan-transcriptome and characterized its transcriptional regulatory landscape in response to cold stress. We performed Iso-Seq and RNA-Seq of 11 rice cultivars subjected to a time-course cold treatment. Our analyses revealed that alternative splicing-regulated gene expression plays a significant role in the cold stress response. Moreover, we identified CATALASE C (OsCATC) and Os03g0701200 as candidate genes for engineering enhanced cold tolerance. Importantly, we uncovered central roles for the two serine-arginine-rich proteins OsRS33 and OsRS2Z38 in cold tolerance. Our analysis of cold tolerance and resequencing data from a diverse collection of 165 rice cultivars suggested that OsRS2Z38 may be a key selection gene in japonica domestication for cold adaptation, associated with the adaptive evolution of rice. This study systematically investigated the distribution, dynamic changes, and regulatory mechanisms of alternative splicing in rice under cold stress. Overall, our work generates a rich resource with broad implications for understanding the genetic basis of cold response mechanisms in plants.

Arabinogalactan proteins (AGPs) are complex proteoglycans present in plant cell walls across the kingdom. They play crucial roles in biological functions throughout the plant life cycle. In this study, we identified 43 gene members of the AG peptide (an AGP subfamily) within the rice genome, detailing their structure, protein-conserved domains, and motif compositions for the first time. We also examined the expression patterns of these genes across 18 tissues and organs, especially the different parts of the flower (anthers, pollen, pistil, sperm cells, and egg cells). Interestingly, the expression of some AG peptides is mainly present in the pollen grain. Transcription data and GUS staining confirmed that OsAGP6P—a member of the AG peptide gene family—is expressed in the stamen during pollen development stages 11–14, which are critical for maturation as microspores form after meiosis of pollen mother cells. It became noticeable from stage 11, when exine formation occurred—specifically at stage 12, when the intine began to develop. The overexpression of this gene in rice decreased the seed-setting rate (from 91.5% to 30.5%) and plant height (by 21.9%) but increased the tillering number (by 34.1%). These results indicate that AGP6P contributes to the development and fertility of pollen, making it a valuable gene target for future genetic manipulation of plant sterility through gene overexpression or editing.