Abstract
BackgroundChloroplast genome information is critical to understanding forms of photosynthesis in the plant kingdom. During the evolutionary process, plants have developed different photosynthetic strategies that are accompanied by complementary biochemical and anatomical features. Members of family Chenopodiaceae have species with C3 photosynthesis, and variations of C4 photosynthesis in which photorespiration is reduced by concentrating CO2 around Rubisco through dual coordinated functioning of dimorphic chloroplasts. Among dicots, the family has the largest number of C4 species, and greatest structural and biochemical diversity in forms of C4 including the canonical dual-cell Kranz anatomy, and the recently identified single cell C4 with the presence of dimorphic chloroplasts separated by a vacuole. This is the first comparative analysis of chloroplast genomes in species representative of photosynthetic types in the family.ResultsMethodology with high throughput sequencing complemented with Sanger sequencing of selected loci provided high quality and complete chloroplast genomes of seven species in the family and one species in the closely related Amaranthaceae family, representing C3, Kranz type C4 and single cell C4 (SSC4) photosynthesis six of the eight chloroplast genomes are new, while two are improved versions of previously published genomes. The depth of coverage obtained using high-throughput sequencing complemented with targeted resequencing of certain loci enabled superior resolution of the border junctions, directionality and repeat region sequences. Comparison of the chloroplast genomes with previously sequenced plastid genomes revealed similar genome organization, gene order and content with a few revisions. High-quality complete chloroplast genome sequences resulted in correcting the orientation the LSC region of the published Bienertia sinuspersici chloroplast genome, identification of stop codons in the rpl23 gene in B. sinuspersici and B. cycloptera, and identifying an instance of IR expansion in the Haloxylon ammodendron inverted repeat sequence. The rare observation of a mitochondria-to-chloroplast inter-organellar gene transfer event was identified in family Chenopodiaceae.ConclusionsThis study reports complete chloroplast genomes from seven Chenopodiaceae and one Amaranthaceae species. The depth of coverage obtained using high-throughput sequencing complemented with targeted resequencing of certain loci enabled superior resolution of the border junctions, directionality, and repeat region sequences. Therefore, the use of high throughput and Sanger sequencing, in a hybrid method, reaffirms to be rapid, efficient, and reliable for chloroplast genome sequencing.
Highlights
Plastids convert light energy into chemical energy and are an essential site for the biosynthesis of pigments, lipids, several amino acids and vitamins [1, 2]
Three large contigs with overlapping 5′ and 3′ regions were generated during genome assembly for A. retroflexus, B. muricata, B. cycloptera, B. sinuspersici, H. ammodendron, S. aralocaspica, and S. maritima
These three contigs were identified as Large Single Copy (LSC), Small Single Copy (SSC), and inverted repeat (IR) via BLAST homology alignment [42], GE-Seq—Annotation of Organellar Genomes [43] and Dual Organellar Genome Annotator (DOGMA) gene identity prediction [44]
Summary
Plastids convert light energy into chemical energy and are an essential site for the biosynthesis of pigments, lipids, several amino acids and vitamins [1, 2]. Chloroplast genome information is critical to understanding forms of photosynthesis in the plant king‐ dom. The family has the largest number of C 4 species, and greatest structural and biochemical diversity in forms of C 4 including the canonical dual-cell Kranz anatomy, and the recently identified single cell C4 with the presence of dimorphic chloroplasts sepa‐ rated by a vacuole. This is the first comparative analysis of chloroplast genomes in species representative of photosyn‐ thetic types in the family
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