Understanding the role of meiosis-specific long intergenic non-coding RNAs in Arabidopsis thaliana

Project Summary

Long non-coding RNAs (lncRNAs) are polyadenylated non protein-coding RNAs that are typically greater than 200 bp long. LncRNAs can be divided into two classes: antisense non-coding RNAs (ancRNAs), which overlap protein-coding genes, and long intergenic non-coding RNAs (lincRNAs), which do not (Nam & Bartel, 2012). There are several lines of evidence that imply these lncRNAs may play roles in meiosis. A collaborator generated RNA-Seq data from isolated Arabidopsis meiocytes which we utilized  to create a pipeline that was used to identify novel meiosis-specific lncRNAs, including 203 lincRNAs. We propose that these lincRNAs may play a critical role in the regulation of Arabidopsismeiosis and are searching for meiotic defects in a T-DNA directed mutant screen.

We used an Alexander stain to determine the viability of pollen, which demonstrated whether meiosis successfully generated functional germ cells in the male haploid cells.  We next used acetocarmine staining to observe tetrads, a structure that contains the four products of one meiosis. Disruption of pollen viability and the ability to form tetrads could indicate a meiotic defect. In addition to these two techniques, we performed chromosome spreads on one specific lincRNA T-DNA line. We visualized these spreads under the microscope to compare phenotypes of mutant meiocytes to those of wild-type. Preliminary data from this specific lincRNA mutant may indicate a defect in meiosis.

My Experience

This was a highly valuable experience for me as a student and a scientist. I learned so much under the tutelage of my mentor, and was grateful to work with someone who had much more experience with scientific research who could patiently teach  and guide me. I gained experience in entirely new techniques, and refreshed other techniques previously performed. This work experience has increased my confidence in my ability to perform research in a lab, and has also fostered a great appreciation for plant science.