A5066335450- Chromosomal and Genetic Variations
- CRISPR and Genetic Engineering
- Plant tissue culture and regeneration
- Plant Molecular Biology Research
- Plant Virus Research Studies
- Photosynthetic Processes and Mechanisms
- Plant Disease Resistance and Genetics
- Genetic Mapping and Diversity in Plants and Animals
- Wheat and Barley Genetics and Pathology
- Botanical Research and Applications
- Plant Reproductive Biology
- Legume Nitrogen Fixing Symbiosis
- Plant nutrient uptake and metabolism
- DNA Repair Mechanisms
- RNA Research and Splicing
- Genomics and Phylogenetic Studies
- Plant Diversity and Evolution
- Garlic and Onion Studies
- Insect symbiosis and bacterial influences
- Antioxidant Activity and Oxidative Stress
- Genomics and Chromatin Dynamics
- Retinal Development and Disorders
- Plant Pathogens and Resistance
- Silicon Effects in Agriculture
- Edible Oils Quality and Analysis
Weizmann Institute of Science
2015-2024
Summary Current breeding relies mostly on random mutagenesis and recombination to generate novel genetic variation. However, targeted genome editing is becoming an increasingly important tool for precise plant breeding. Using the CRISPR ‐Cas system combined with bean yellow dwarf virus rolling circle replicon, we optimized a method gene replacement in tomato. The carotenoid isomerase ( CRTISO ) phytoene synthase 1 PSY genes from biosynthesis pathway were chosen as targets due their easily...
Abstract Speciation via interspecific or intergeneric hybridization and polyploidization triggers genomic responses involving genetic epigenetic alterations. Such modifications may be induced by small RNAs, which affect key cellular processes, including gene expression, chromatin structure, cytosine methylation transposable element (TE) activity. To date, the role of RNAs in context wide has received little attention. In this work, we performed high-throughput sequencing parental, hybrid,...
Gene targeting, which is homologous recombination-mediated integration of an extra-chromosomal DNA segment into a chromosomal target sequence, enables the precise disruption or replacement any gene. Despite its value as molecular genetic tool, gene targeting remains inefficient technology in most species. We report that expression yeast RAD54 gene, member SWI2 / SNF2 chromatin remodeling family, enhances Arabidopsis by one to two orders magnitude, from 10 -4 -3 WT plants -2 -1 . show events,...
Meiotic recombination is tightly regulated by cis - and trans -acting factors. Although DNA methylation chromatin remodeling affect chromosome structure, their impact on meiotic not well understood. To study the effect of landscape chromosomal recombination, we analyzed in decreased 1 ( ddm1 ) mutant. DDM1 a SWI2/SNF2-like chromatin-remodeling protein necessary for heterochromatin maintenance Arabidopsis thaliana . The rate between markers located euchromatic regions was significantly higher...
The rate of crossover, the reciprocal exchanges homologous chromosomal segments, is not uniform along chromosomes differing between male and female meiocytes. To better understand factors regulating this variable landscape, we performed a detailed genetic epigenetic analysis 737 crossover events in Arabidopsis thaliana. Crossovers were more frequent than expected promoters. Three DNA motifs enriched regions less abundant crossover-poor pericentric identified. One these motifs, CCN repeat,...
Many plant roots acquire inorganic phosphate (Pi) from soils directly through the root-soil interface via high-affinity Pi transporters and/or symbiotic associations between cortical cells and arbuscular mycorrhizal fungi. In tomato, three (LePT3, LePT4, LePT5) are up-regulated upon colonization by this study, role of LePT4 in tomato is elucidated molecular physiological characterizations a loss-of-function mutant lept4. absence infection under solution-Pi concentrations (Cp) 0.05 mM 0.5 mM,...
Abstract CRISPR/Cas9 is widely used for precise mutagenesis through targeted DNA double-strand breaks (DSBs) induction followed by error-prone repair. A better understanding of this process requires measuring the rates cutting, error-prone, and repair, which have remained elusive so far. Here, we present a molecular computational toolkit multiplexed quantification DSB intermediates repair products single-molecule sequencing. Using approach, characterize dynamics induction, processing at...
Abstract RADiation sensitive52 (RAD52) mediates RAD51 loading onto single-stranded DNA ends, thereby initiating homologous recombination and catalyzing annealing. RAD52 is highly conserved among eukaryotes, including animals fungi. This article reports that homologs are present in all plants whose genomes have undergone extensive sequencing. Computational analyses suggest a very early gene duplication, followed by later lineage-specific duplications, during the evolution of higher plants....
Targeted modification of the genome is an important genetic tool, which can be achieved via homologous, non-homologous or site-specific recombination. Although numerous efforts have been made, such a tool does not exist for routine applications in plants. This work describes simple and useful method targeted mutagenesis gene targeting, tailored to floral-dip transformation Arabidopsis, by means specific protein expression egg cell. Proteins stably transiently expressed under...
DNA double-stranded breaks (DSBs) generated by the Cas9 nuclease are commonly repaired via nonhomologous end-joining (NHEJ) or homologous recombination (HR). However, little is known about unrepaired DSBs and type of damage they trigger in plants. We designed an assay that detects loss heterozygosity (LOH) somatic cells, enabling study a broad range DSB-induced genomic events. The system relies on mapped phenotypic marker which produces light purple color (betalain pigment) all plant...
Abstract In higher plants, the plastidial NADH dehydrogenase (Ndh) complex supports nonphotochemical electron fluxes from stromal donors to plastoquinones. Ndh functions in chloroplasts are not clearly established; however, its activity was linked prevention of overreduction stroma, especially under stress conditions. Here, we show by characterization OrrDs, a dominant transposon-tagged tomato (Solanum lycopersicum) mutant deficient NDH-M subunit, that this is also essential for fruit...
Homologous recombination (HR) in somatic cells is not as well understood meiotic and thought to be rare. In a previous study, we showed that Inter-Homologous Somatic Recombination (IHSR) can achieved by targeted induction of DNA double-strand breaks (DSBs). Here, designed novel IHSR assay investigate this phenomenon greater depth. We utilized F1 hybrids from divergent parental lines, each with different mutation at the Carotenoid isomerase (CRTISO) locus. events, namely crossover or gene...
Summary Meiotic recombination is a fundamental biological process that plays central role in the evolution and breeding of plants. We have developed new seed‐based assay for meiotic Arabidopsis. The based on transformation green red fluorescent markers expressed under seed‐specific promoter. A total 74 T‐DNA were isolated, sequenced mapped both physically genetically. Lines containing map 1–20 cM apart crossed to produce tester lines with two linked cis yielding seeds fluoresced green. show...
Starches extracted from most plant species are phosphorylated. alpha-Glucan water dikinase (GWD) is a key enzyme that controls the phosphate content of starch. In absence its activity starch degradation impaired, leading to excess phenotype in Arabidopsis and potato leaves, reduced cold sweetening tubers. Here, we characterized transposon insertion (legwd::Ds) tomato GWD (LeGWD) gene caused male gametophytic lethality. The mutant pollen had was associated with reduction germination. SEM TEM...
Agrobacterium tumefaciens mediated T-DNA integration is a common tool for plant genome manipulation. However, there controversy regarding whether biased towards genes or randomly distributed throughout the genome. In order to address this question, we performed high-throughput mapping of T-DNA-genome junctions obtained in absence selection at several time points after infection. were detected as early 6 hours post-infection. distribution was apparently uniform chromosomes, yet local biases...
Meiotic recombination is the main driver of genetic diversity in wheat breeding. The rate and location crossover (CO) events are regulated by epigenetic factors. In wheat, most COs occur subtelomeric regions but rare centromeric pericentric areas. aim this work was to increase both “hot” “cold” chromosomal locations. We used Virus-Induced gene Silencing (VIGS) downregulate expression recombination-suppressing genes XRCC2 FANCM maintenance MET1 DDM1 during meiosis. VIGS suppresses a dominant,...
Homologous recombination (HR) typically occurs during meiosis between homologs, at a few unplanned locations along the chromosomes. In this study, we tested whether targeted homologous chromosomes can be achieved via Clustered Regulatory Interspaced Short Palindromic Repeat associated protein Cas9 (CRISPR-Cas9)-induced DNA double-strand break (DSB) repair in Arabidopsis thaliana. Our experimental system includes targets for DSB induction euchromatic and heterochromatic genomic regions of...
Gene Targeting (GT) is the integration of an introduced vector into a specific chromosomal site, via homologous recombination. It considered effective tool for precise genome editing, with far-reaching implications in biological research and biotechnology, widely used mice, potential becoming routine many species. Nevertheless, epigenetic status targeted allele remains largely unexplored. Using GT-modified lines model plant Arabidopsis thaliana, we show that DNA methylation profile locus...