High-throughput discovery of plastid genes causing albino phenotypes in ornamental chimeric plants

Chimeric plants composed of green and albino tissues have great ornamental value.To unveil the functional genes responsible for albino phenotypes in chimeric plants,we inspected the complete plastid genomes(plastomes)in green and albino leaf tissues from 23 ornamental chimeric plants belonging to 20 species,including monocots,dicots,and gymnosperms.In nine chimeric plants,plastomes were identical between green and albino tissues.Meanwhile,another 14 chimeric plants were heteroplasmic,showing a mutation between green and albino tissues.We identified 14 different point mutations in eight functional plastid genes related to plastid-encoded RNA polymerase(rpo)or photosystems which caused albinism in the chimeric plants.Among them,12 were deleterious mutations in the target genes,in which early termination appeared due to small deletion-mediated frameshift or single nucleotide substitution.Another was single nucleotide substitution in an intron of the ycf3 and the other was a missense mutation in coding region of the rpoC2 gene.We inspected chlorophyll structure,protein functional model of the rpoC2,and expression levels of the related genes in green and albino tissues of Reynoutria japonica.A single amino acid change,histidine-to-proline substitution,in the rpoC2 protein may destabilize the peripheral helix of plastid-encoded RNA polymerase,impairing the biosynthesis of the photosynthesis system in the albino tissue of R.japonica chimera plant.

Nuclear and chloroplast genome diversity revealed by low-coverage whole-genome shotgun sequence in 44 Brassica oleracea breeding lines

Whole-genome shogun sequence(WGS)data generated by next-generation sequencing(NGS)platforms are a valuable resource for crop improvement.We produced 5–6×WGS coverage of 44 Brassica oleracea breeding lines representing seven subspecies/morphotypes:cabbage,broccoli,cauliflower,kailan,kale,Brussels sprout,and kohlrabi to systematically evaluate the nuclear and chloroplast(Cp)diversity in the 44 B.oleracea breeding lines.We then exploited the impact of low-coverage NGS by evaluating nuclear genome diversity and assembly,annotation of complete chloroplast(Cp)genomes and 45 S nuclear ribosomal DNA(45S nrDNA)sequences,and copy number variation for major repeats.Nuclear genome diversity analysis has revealed a total of 496463 SNPs and 37493 indels in the nuclear genome across the 44 accessions.Interestingly,some SNPs showed subspecies enrichment at certain chromosomal regions.The assembly of complete Cp genomes contained 153361–153372 bp with 37 variants including SNPs and indels.The 45S nrDNA transcription unit was 5802 bp long with a total of 31 SNPs from the 44 lines.The phylogenetic tree inferred from the nuclear and Cp genomes coincided and clustered broccoli,cauliflower,and kailan in one group and cabbage,Brussels sprout,kale,and kohlrabi in another group.The morphotypes diverged during the last 0.17 million years.The Cp genome diversity reflected the unique cytoplasm of each subspecies,and revealed that the cytoplasm of many breeding lines was replaced and intermingled via inter-subspecies crosses during the breeding process instead.The polymorphic Cp markers provide a classification system for the cytoplasm types in B.oleracea.Furthermore,copy numbers of major transposable elements(TEs)showed high diversity among the 44 accessions,indicating that many TEs have become active recently.Overall,we demonstrated a comprehensive utilization of low-coverage NGS data and might shed light on the genetic diversity and evolution of diverse B.oleracea morphotypes.