The class 2/type II clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system has been used successfully for simultaneous modification of multiple loci in plants. Two general strategies have been applied to coexpressmultiple single guide RNAs (sgRNAs) to achieve multiplex gene editing in plant cells. One is to construct the multiple guide RNA expression cassettes into separa...

Just as Koch's postulates formed the foundation of early infectious disease study, Stanley Falkow's molecular Koch's postulates define best practice in determining whether a specific gene contributes to virulence of a pathogen. Fundamentally, these molecular postulates state that if a gene is involved in virulence, its removal will compromise virulence. Likewise, its reintroduction should resto...

Protoplasts are cells without cell walls. In plants protoplasts are usually liberated by tissue digestion with cellulases and pectinases purified from wood-rotting fungi. To keep the plasma membrane from bursting during protoplast isolation, an isotonic or slightly hypertonic medium is used. Because of the lack of external constraints on the membrane, protoplasts are spherical (figure 1B). One ...

Ferns (Pteridophyta) are very important members of the plant kingdom that lag behind other taxa with regards to our understanding of their genetics, genomics, andmolecular biology. We report here, to our knowledge, the first instance of stable transformation of fern with recovery of transgenic sporophytes. Spores of the arsenic hyperaccumulating fern Pteris vittata and tetraploid ‘C-fern Expres...

Modern transformation and genome editing techniques have shown great success across a broad variety of organisms. However, no study of successfully applied genome editing has been reported in a dinoflagellate despite the first genetic transformation of Symbiodinium being published about 20 years ago. Using an array of different available transformation techniques, we attempted to transform Symb...

A full-length cDNA clone of the potato virus Y (PVY) genome was obtained after cloning difficulties in Escherichia coli were overcome. These difficulties were mainly due to the expression of the CI gene from upstream prokaryotic promoter-like elements within the PVY genome. To overcome this problem, PVY cDNA was maintained in two subclones which were ligated before infection. A plasmid in which...

In this paper, computational fluid dynamics (CFD) is utilized to characterize a prototype biolistic delivery system, the biomedical device based on the contoured-shock-tube design (CST), with the aim at investigating shocks induced flow instabilities within the contoured shock tube. The shock/interface interactions, the growth of perturbation at an interface between two fluids of different dens...

Since yeast is amenable to mitochondrial transformation, designed mutations can be introduced in the mitochondrially encoded subunits of the respiratory complexes. In the present work, six mutations have been introduced by the biolistic method into yeast (Saccharomyces cerevisiae) cytochrome oxidase subunits I and III. The effects of these mutations on respiratory growth competence, cytochrome ...

We investigated the size of flanking DNA incorporated into the tobacco plastid genome alongside a selectable antibiotic resistance mutation. The results showed that integration of a long uninterrupted region of homologous DNA, rather than of small fragments as previously thought, is the more likely event in plastid transformation of land plants. Transforming plasmid pJS75 contains a 6.2-kb DNA ...

Mutations have been inserted into the virion and complementary sense ORFs encoding proteins with M(r)s in excess of 9 kDa of both DNA A and DNA B of potato yellow mosaic geminivirus (PYMV). Wild-type and mutant monomeric clones were tested for their ability to replicate, produce PYMV-specific DNA, spread and cause symptoms in Nicotiana benthamiana plants following biolistic inoculation. Dimeric...