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Updated: Feb 5, 2026

Standardized Method for High-throughput Sterilization of Arabidopsis Seeds

Standardized Method for High-throughput Sterilization of Arabidopsis Seeds

Published on: October 17, 2017

Arabidopsis

Nancy R Forsthoefel¹, Kendra A Klag¹, Savannah R McNichol¹

¹Program in Biochemistry, Biophysics, and Molecular Biology, Whitman College, Walla Walla, Washington 99362.

Plant Physiology

|September 13, 2018

Summary

This summary is machine-generated.

Plant intracellular Ras-group leucine-rich repeat (LRR) proteins (PIRLs) are crucial for gametogenesis. PIRL6 is essential for male and female gametophyte development, with its expression regulated by alternative splicing in sporophytes.

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Lateral Root Inducible System in Arabidopsis and Maize

Lateral Root Inducible System in Arabidopsis and Maize

Published on: January 14, 2016

Area of Science:

Plant molecular biology
Developmental genetics
Arabidopsis research

Background:

Plant intracellular Ras-group leucine-rich repeat (LRR) proteins (PIRLs) are involved in cell signaling.
Some PIRL family members in Arabidopsis thaliana are implicated in pollen development.
PIRL6 has no known knockout alleles, suggesting essential gametophyte functions.

Purpose of the Study:

To investigate the expression and function of PIRL6 in Arabidopsis gametogenesis.
To determine if PIRL6 plays a role in both male and female gametophyte development.
To explore the regulatory mechanisms controlling PIRL6 expression.

Main Methods:

RNA interference (RNAi) to induce PIRL6 knockdown.
Analysis of gametogenesis defects in knockdown mutants.
Expression analysis using wild-type and sporocyteless (spl) mutant flowers.
PIRL6-GFP fusion construct for expression localization.
cDNA sequencing and analysis of alternative splicing.
Nonsense-mediated decay (NMD) assays using upf3 mutants.

Main Results:

PIRL6 knockdown caused defects in pollen development and embryo sac formation.
PIRL6 is expressed in gametophytes, confirmed by expression in wild-type but not spl mutant flowers.
PIRL6-GFP localized to pollen and embryo sac.
Multiple alternative, non-functional PIRL6 mRNAs containing introns and premature termination codons were detected.
These aberrant mRNAs accumulate in upf3 mutants, indicating they are NMD targets.
Alternative splicing negatively regulates functional PIRL6 expression in sporophyte tissues.

Conclusions:

PIRL6 is essential for both male and female gametogenesis in Arabidopsis.
Sporophytic expression of PIRL6 is negatively regulated by unproductive alternative splicing.
This posttranscriptional mechanism likely minimizes PIRL6 protein in sporophytes, allowing transcription of an overlapping adjacent gene.