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Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy
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Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy

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The 5' stem-loop and its role in mRNA stability in maize S cytoplasmic male sterility

Hailin Xiao1, Fangdong Zhang, Yonglian Zheng

  • 1State Key Laboratory of Crop Genetic Improvement and National Center of Crop Molecular Breeding, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.

The Plant Journal : for Cell and Molecular Biology
|September 12, 2006
PubMed

Insights

A 5' stem-loop deletion in maize mitochondrial transcripts causes degradation, leading to reduced fertility restoration in cytoplasmic male sterility (CMS-S) plants. This explains lower transcript levels in restored microspores.

Area of Science:

  • Plant Molecular Biology
  • Mitochondrial Genetics
  • Maize Breeding

Background:

  • The orf355-orf77 region of the maize mitochondrial genome is linked to S cytoplasmic male sterility (CMS-S).
  • Transcripts from this region are significantly reduced in fertility-restored microspores compared to sterile plants.

Purpose of the Study:

  • To investigate the molecular mechanism behind the reduction of orf355-orf77 transcripts in fertility-restored maize.
  • To analyze the 5' and 3' termini of the 1.6-kb and 2.8-kb transcripts associated with CMS-S.

Main Methods:

  • 3' RACE (Rapid Amplification of cDNA Ends) analysis to map polyadenylation sites.
  • Primer extension to analyze the 5' terminus of the 1.6-kb transcript.

Main Results:

  • Polyadenylation sites were located near a 3' stem-loop structure in both sterile and restored microspores, with no significant differences.
  • A nine-nucleotide deletion was identified at the 5' terminus of the 1.6-kb transcript exclusively in fertility-restored microspores.
  • This deletion eliminated a crucial 5' stem-loop structure, potentially leading to transcript degradation.

Conclusions:

  • The elimination of the 5' stem-loop in fertility-restored microspores is proposed as the cause for the degradation of the 1.6-kb transcript.
  • The reduction in the 2.8-kb transcript, which can be cleaved to produce the 1.6-kb transcript, is a consequence of this degradation pathway.

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