14 research outputs found
Wolfberry genomes and the evolution of Lycium (Solanaceae)
AbstractWolfberry Lycium, an economically important genus of the Solanaceae family, contains approximately 80 species and shows a fragmented distribution pattern among the Northern and Southern Hemispheres. Although several herbaceous species of Solanaceae have been subjected to genome sequencing, thus far, no genome sequences of woody representatives have been available. Here, we sequenced the genomes of 13 perennial woody species of Lycium, with a focus on Lycium barbarum. Integration with other genomes provides clear evidence supporting a whole-genome triplication (WGT) event shared by all hitherto sequenced solanaceous plants, which occurred shortly after the divergence of Solanaceae and Convolvulaceae. We identified new gene families and gene family expansions and contractions that first appeared in Solanaceae. Based on the identification of self-incompatibility related-gene families, we inferred that hybridization hotspots are enriched for genes that might be functioning in gametophytic self-incompatibility pathways in wolfberry. Extremely low expression of LOCULE NUBER (LC) and COLORLESS NON-RIPENING (CNR) orthologous genes during Lycium fruit development and ripening processes suggests functional diversification of these two genes between Lycium and tomato. The existence of additional flowering locus C-like MADS-box genes might correlate with the perennial flowering cycle of Lycium. Differential gene expression involved in the lignin biosynthetic pathway between Lycium and tomato likely illustrates woody and herbaceous differentiation. We also provide evidence that Lycium migrated from Africa into Asia, and subsequently from Asia into North America. Our results provide functional insights into Solanaceae origins, evolution and diversification.</jats:p
Switchable Ferro‑, Ferri‑, and Antiferromagnetic States in a Piezo- and Hydrochromic Metal–Organic Framework
The MnÂ(II) metal–organic
framework with a viologen-based tetracarboxylate ligand exhibits reversible
optical (color) and magnetic changes concomitant with stimuli-induced
electron transfer from carboxylate to viologen. Compression causes
a magnetic transformation from ferro- to ferrimagnetic, while water
release/reuptake switches the magnetic behavior between ferro- and
antiferromagnetic
Wolfberry genomes and the evolution of Lycium (Solanaceae)
Wolfberry Lycium, an economically important genus of the Solanaceae family, contains
approximately 80 species and shows a fragmented distribution pattern among the Northern
and Southern Hemispheres. Although several herbaceous species of Solanaceae have been
subjected to genome sequencing, thus far, no genome sequences of woody representatives
have been available. Here, we sequenced the genomes of 13 perennial woody species of
Lycium, with a focus on Lycium barbarum. Integration with other genomes provides clear
evidence supporting a whole-genome triplication (WGT) event shared by all hitherto
sequenced solanaceous plants, which occurred shortly after the divergence of Solanaceae
and Convolvulaceae. We identified new gene families and gene family expansions and
contractions that first appeared in Solanaceae. Based on the identification of selfincompatibility related-gene families, we inferred that hybridization hotspots are enriched
for genes that might be functioning in gametophytic self-incompatibility pathways in wolfberry. Extremely low expression of LOCULE NUBER (LC) and COLORLESS NON-RIPENING
(CNR) orthologous genes during Lycium fruit development and ripening processes suggests
functional diversification of these two genes between Lycium and tomato. The existence of
additional flowering locus C-like MADS-box genes might correlate with the perennial flowering
cycle of Lycium. Differential gene expression involved in the lignin biosynthetic pathway
between Lycium and tomato likely illustrates woody and herbaceous differentiation. We also
provide evidence that Lycium migrated from Africa into Asia, and subsequently from Asia into
North America. Our results provide functional insights into Solanaceae origins, evolution and
diversification.https://www.nature.com/commsbiopm2021BiochemistryGeneticsMicrobiology and Plant Patholog
“Perfect” designer chromosome V and behavior of a ring derivative
INTRODUCTION
The
Saccharomyces cerevisiae
2.0 project (Sc2.0) aims to modify the yeast genome with a series of densely spaced designer changes. Both a synthetic yeast chromosome arm (synIXR) and the entirely synthetic chromosome (synIII) function with high fitness in yeast. For designer genome synthesis projects, precise engineering of the physical sequence to match the specified design is important for the systematic evaluation of underlying design principles. Yeast can maintain nuclear chromosomes as rings, occurring by chance at repeated sequences, although the cyclized format is unfavorable in meiosis given the possibility of dicentric chromosome formation from meiotic recombination. Here, we describe the de novo synthesis of synthetic yeast chromosome V (synV) in the “Build-A-Genome China” course, perfectly matching the designer sequence and bearing loxPsym sites, distinguishable watermarks, and all the other features of the synthetic genome. We generated a ring synV derivative with user-specified cyclization coordinates and characterized its performance in mitosis and meiosis.
RATIONALE
Systematic evaluation of underlying Sc2.0 design principles requires that the final assembled synthetic genome perfectly match the designed sequence. Given the size of yeast chromosomes, synthetic chromosome construction is performed iteratively, and new mutations and unpredictable events may occur during synthesis; even a very small number of unintentional nucleotide changes across the genome could have substantial effects on phenotype. Therefore, precisely matching the physical sequence to the designed sequence is crucial for verification of the design principles in genome synthesis. Ring chromosomes can extend those design principles to provide a model for genomic rearrangement, ring chromosome evolution, and human ring chromosome disorders.
RESULTS
We chemically synthesized, assembled, and incorporated designer chromosome synV (536,024 base pairs) of
S. cerevisiae
according to Sc2.0 principles, based on the complete nucleotide sequence of native yeast chromosome V (576,874 base pairs). This work was performed as part of the “Build-A-Genome China” course in Tianjin University.
We corrected all mutations found—including duplications, substitutions, and indels—in the initial synV strain by using integrative cotransformation of the precise desired changes and by means of a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)–based method. Altogether, 3331 corrected base pairs were required to match to the designed sequence. We generated a strain that exactly matches all designer sequence changes that displays high fitness under a variety of culture conditions. All corrections were verified with whole-genome sequencing; RNA sequencing revealed only minor changes in gene expression—most notably, decreases in expression of genes relocated near synthetic telomeres as a result of design.
We constructed a functional circular synV (ring_synV) derivative in yeast by precisely joining both chromosome ends (telomeres) at specified coordinates. The ring chromosome showed restoration of subtelomeric gene expression levels. The ring_synV strain exhibited fitness comparable with that of the linear synV strain, revealed no change in sporulation frequency, but notably reduced spore viability. In meiosis, heterozygous or homozygous diploid ring_wtV and ring_synV chromosomes behaved similarly, exhibiting substantially higher frequency of the formation of zero-spore tetrads, a type that was not seen in the rod chromosome diploids. Rod synV chromosomes went through meiosis with high spore viability, despite no effort having been made to preserve meiotic competency in the design of synV.
CONCLUSION
The perfect designer-matched synthetic chromosome V provides strategies to edit sequence variants and correct unpredictable events, such as off-target integration of extra copies of synthetic DNA elsewhere in the genome. We also constructed a ring synthetic chromosome derivative and evaluated its fitness and stability in yeast. Both synV and synVI can be circularized and can power yeast cell growth without affecting fitness when gene content is maintained. These fitness and stability phenotypes of the ring synthetic chromosome in yeast provide a model system with which to probe the mechanism of human ring chromosome disorders.
Synthesis, cyclization, and characterization of
synV
.
(
A
) Synthetic chromosome V (synV, 536,024 base pairs) was designed in silico from native chromosome V (wtV, 576,874 base pairs), with extensive genotype modification designed to be phenotypically neutral. (
B
) CRISPR/Cas9 strategy for multiplex repair. (
C
) Colonies of wtV, synV, and ring_synV strains.
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