Capturing the biofuel wellhead and powerhouse: the chloroplast and mitochondrial genomes of the leguminous feedstock tree Pongamia pinnata

PLoS One. 2012;7(12):e51687. doi: 10.1371/journal.pone.0051687. Epub 2012 Dec 14.

Abstract

Pongamia pinnata (syn. Millettia pinnata) is a novel, fast-growing arboreal legume that bears prolific quantities of oil-rich seeds suitable for the production of biodiesel and aviation biofuel. Here, we have used Illumina® 'Second Generation DNA Sequencing (2GS)' and a new short-read de novo assembler, SaSSY, to assemble and annotate the Pongamia chloroplast (152,968 bp; cpDNA) and mitochondrial (425,718 bp; mtDNA) genomes. We also show that SaSSY can be used to accurately assemble 2GS data, by re-assembling the Lotus japonicus cpDNA and in the process assemble its mtDNA (380,861 bp). The Pongamia cpDNA contains 77 unique protein-coding genes and is almost 60% gene-dense. It contains a 50 kb inversion common to other legumes, as well as a novel 6.5 kb inversion that is responsible for the non-disruptive, re-orientation of five protein-coding genes. Additionally, two copies of an inverted repeat firmly place the species outside the subclade of the Fabaceae lacking the inverted repeat. The Pongamia and L. japonicus mtDNA contain just 33 and 31 unique protein-coding genes, respectively, and like other angiosperm mtDNA, have expanded intergenic and multiple repeat regions. Through comparative analysis with Vigna radiata we measured the average synonymous and non-synonymous divergence of all three legume mitochondrial (1.59% and 2.40%, respectively) and chloroplast (8.37% and 8.99%, respectively) protein-coding genes. Finally, we explored the relatedness of Pongamia within the Fabaceae and showed the utility of the organellar genome sequences by mapping transcriptomic data to identify up- and down-regulated stress-responsive gene candidates and confirm in silico predicted RNA editing sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Codon
  • Fabaceae / genetics
  • Fabaceae / metabolism
  • Gene Order
  • Genome, Chloroplast*
  • Genome, Mitochondrial*
  • Millettia / classification
  • Millettia / genetics*
  • Millettia / metabolism
  • Open Reading Frames
  • Phylogeny
  • Plastids / genetics
  • RNA Editing
  • RNA, Plant

Substances

  • Codon
  • RNA, Plant

Grants and funding

The authors thank BioEnergy Plantation Australia for research support in Pongamia, BES Pty Ltd. for university liaison. Research was also funded by an Australian Research Council Centre of Excellence grant to PMG. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.