Project description:EMS-mutagenized D. melanogaster populations with 4xbicoid were sequenced at the 1st, 3rd, 7th, 9th and 15th generation to analyze genomic changes during experimental evolution. Non-mutagenized populations were sequenced in parallel, representing standing variations.
Project description:The rdr6.1 missense allele was found by TILLING in EMS mutagenized plants of Medicago truncatula. To analyse the impact of this point mutation on small RNA populations, we sequenced small RNA from two sibling lines carrying the mutation and the corresponding wild-type plants.
Project description:NaCl-resistant Saccharomyces cerevisiae mutant was obtained by evolutionary engineering. EMS mutagenized culture was used as the initial population for the selection procedure. Gradually increasing levels of NaCl stress was applied through 40 successive batch cultivations. The reference strain could not grow even at 0.85 M NaCl whereas this mutant was shown to be resistant up to 1.45 M NaCl concentration. Whole-genome microarray analysis was used to identify the NaCl resistance mechanisms by comparing NaCl-resistant mutant strain and the reference strain in the absence of NaCl stress.
Project description:The rdr6.1 missense allele was found by TILLING in EMS mutagenized plants of Medicago truncatula. To analyse the impact of this point mutation on small RNA populations, we sequenced small RNA from two sibling lines carrying the mutation and the corresponding wild-type plants. libraries from two sibling lines carrying the rdr6.1 mutation and the corresponding wild-type plants
Project description:<p>Pigmented rice (<em>Oryza sativa L.</em>) is a rich source of nutrients, but pigmented lines typically have long life cycles and limited productivity. Here we generated genome assemblies of 5 pigmented rice varieties and evaluated the genetic variation among 51 pigmented rice varieties by resequencing an additional 46 varieties. Phylogenetic analyses divided the pigmented varieties into four varietal groups: Geng-japonica, Xian-indica, circum-Aus and circum-Basmati. Metabolomics and ionomics profiling revealed that black rice varieties are rich in aromatic secondary metabolites. We established a regeneration and transformation system and used CRISPR-Cas9 to knock out three flowering time repressors (Hd2, Hd4 and Hd5) in the black Indonesian rice Cempo Ireng, resulting in an early maturing variety with shorter stature. Our study thus provides a multi-omics resource for understanding and improving Asian pigmented rice.</p>
Project description:To investigate the functions of TRANSPARENT TESTA 19 (TT19) on anthocyanin accumulation and stabilization using forward genetic screening approach. EMS mutagenized tt19 mutant to screen tt19 mutant suppressors that could restore the anthocyanin accumulation and the causative mutations were identified as point mutations in the RDR6/SGS3/DCL4 siRNA biogenesis pathway using whole genome re-sequencing. The idea for RNA-Seq and small RNA-Seq analysis is to illustrate the connection between siRNA biogenesis, TT19, and anthocyanin restoration. RNA's of 4 Arabidopsis lines (col-0, tt19 and two TT19 suppressors, all in triplicate) which were under AIC (3% sucrose shaking four days under 24h light) for four days was extracted. Small RNA libraries were prepared by BGI company and sequenced using BGISEQ-500 as 50SE.
Project description:Eukaryotic genomes are heavily regulated by epigenetic marks that often act to modulate the transcriptional control of genetic elements. In Arabidopsis thaliana the ATXR5 and ATXR6 histone methyltransferases, and their cognate H3K27 monomethylation mark, act in transcriptional silencing while also maintaining genome stability by preventing generation of excess DNA corresponding to pericentromeric heterochromatin. In this study we characterize the atxr5 atxr6 transcriptome and its relationship to the DNA damage response which suggests that the atxr5 atxr6 transcriptional defects may be epistatic to the genome instability defects in the mutants. In addition we isolate several factors that modulate both the transcriptional and genomic instability phenotypes of atxr5 atxr6 mutants, which suggest a mechanism for atxr5 atxr6-induced extra DNA involving conflicts between the replicative and transcriptional processes in the cell. PolyA RNA sequencing (RNA-seq), whole-genome resequencing (DNA-seq), and whole-genome bisulfite sequencing (methyl-seq) was performed on Arabidospsis thaliana mutant and wildtype plants. DNA-seq was used to characterize DNA copy number and map EMS-induced mutations, RNA-seq was used to quantify transcript abundance and map EMS-induced mutations, and methyl-seq was used to assess DNA methylation. Details of the relationship between samples in this series and figures in the associated manuscript can be found in Supplemental Table 4 of the associated manuscript. Unless otherwise noted in the description all lines are ecotype Columbia, and all genotypes should be assumed homozygous unless otherwise indicated with a '/'.