Gene Discovery in Reproduction: RNA-seq Analysis of Isolated Pistils from Apomictic and Sexual Eragrostis curvula

Abstract

This chapter describes a comprehensive approach to analyze the differential gene expression of isolated pistils at distinct developmental stages from sexual and apomictic Eragrostis curvula. It integrates the development of reproductive calendars, methods for pistil isolation, and techniques for extracting high-quality RNA from limited tissue samples. This methodology includes detailed steps for integrating sequence-based bioinformatics pipelines and clustering tools to analyze the expression patterns obtained from high-throughput sequencing.Additionally, the methodological outline explains the bioinformatic pipelines used to analyze gene expression patterns, providing insights into molecular mechanisms underlying apomixis. This approach addresses key challenges such as small tissue size and mixed developmental stages by offering a reliable framework for transcriptomic studies in E. curvula.