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Related Concept Videos

Export of Mitochondrial and Chloroplast Genes02:19

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A eukaryotic cell can have up to three different types of genetic systems: nuclear, mitochondrial, and chloroplast. During evolution, organelles have exported many genes to the nucleus; this transfer is still ongoing in some plant species. Approximately 18% of the Arabidopsis thaliana nuclear genome is thought to be derived from the chloroplast’s cyanobacterial ancestor, and around 75% of the yeast genome derived from the mitochondria’s bacterial ancestor. This export has occurred...
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Related Experiment Video

Updated: Apr 23, 2026

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing
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Effective extraction and assembly methods for simultaneously obtaining plastid and mitochondrial genomes.

Wanjun Hao1, Shihang Fan2, Wei Hua1

  • 1Key Laboratory for Biological Sciences of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agriculture Sciences, Wuhan, China.

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|September 25, 2014
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Summary

This study presents a new, cost-effective method for simultaneously sequencing plant plastid and mitochondrial genomes. The approach simplifies DNA extraction and assembly, yielding high-quality results for plant cytoplasmic genome research.

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Area of Science:

  • Plant genomics
  • Molecular biology
  • Bioinformatics

Background:

  • Conventional methods for sequencing plastid DNA (ptDNA) and mitochondrial DNA (mtDNA) require separate preparation, which is challenging due to difficulties in extracting pure DNA from plant tissues.
  • High-throughput sequencing of total DNA for organelle genomes generates massive datasets, leading to high computational costs and potential assembly inaccuracies from pollution reads.
  • A need exists for an efficient, universal, and cost-effective method for simultaneous ptDNA and mtDNA sequencing from plant tissues.

Purpose of the Study:

  • To develop a novel, integrated method for the simultaneous extraction, evaluation, sequencing, and assembly of plant plastid and mitochondrial genomes.
  • To provide a low-cost, time-saving, and reproducible technique suitable for all plant species.
  • To improve the accuracy and efficiency of organelle genome sequencing.

Main Methods:

  • High-quality DNA extraction using Partial Concentration Extraction.
  • DNA purity and dataset size assessment via Vector Control Quantitative Analysis.
  • Paired-end sequencing on a high-throughput platform followed by Two-step Assembly for scaffold generation.
  • Gap-filling and validation using PCR and Sanger sequencing to obtain complete organelle genomes.

Main Results:

  • Successful simultaneous acquisition of complete plastid and mitochondrial genomes.
  • Obtained complete plastid genome of 153,533 nt and mitochondrial genome of 223,412 nt.
  • The method demonstrated high accuracy, confirmed by PCR and Sanger sequencing validation.

Conclusions:

  • A simple, inexpensive, and reproducible method for simultaneous plastid and mitochondrial genome sequencing has been established.
  • The developed technique is time-saving and yields high-quality sequence data.
  • This method is broadly applicable to various plant species, supporting future research on plant cytoplasmic genomes.