Transient assay systems using protoplasts have been utilized in several plant species and are a powerful tool for rapid functional gene analysis and biochemical manipulations. A protoplast system has not been used in switchgrass (Panicum virgatum L.), even though it is a bioenergy crop that has received considerable attention. Here we report the first protocol to isolate large numbers of viable...

Pathogens can colonize all plant organs and tissues. To prevent this, each cell must be capable of autonomously triggering defence. Therefore, it is generally assumed that primary sensors of the immune system are constitutively present. One major primary sensor against bacterial infection is the flagellin sensing 2 (FLS2) pattern recognition receptor (PRR). To gain insights into its expression ...

ABSTRACT Aflatoxin biosynthesis was induced by compounds in filtrates (EF) obtained from cultures consisting of ground maize kernels colonized by Aspergillus flavus. The inducing activity increased to a maximum at 4 days of incubation and then decreased. Amylase activity was detected in the EF, suggesting that the inducers are products of starch degradation (glucose, maltose, and maltotriose). ...

An efficient variety-independent method for producing transgenic eggplants (Solanum melongena L.) via Agrobacterium tumefaciens-mediated genetic transformation was developed. Two weeks old cotyledon explants were transformed by co-cultivation with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pVDH65 carrying the reporter gene β-glucuronidase intron (GUS-INT) and the marker ...

A tropical endophytic isolate of the fungus Fusarium verticillioides (synonym Fusarium moniliforme) obtained from Zea mays was co-transformed with plasmid pNH24 containing the Fusarium oxysporum nitrate reductase nia gene and plasmid pNOM 102 carrying the Escherichia coli β-glucuronidase gusA gene. Transformation frequency for the nia marker was 75 transformants μg vector DNA and introduction o...

Major Accomplishments to date: 1. Genetic Manipulations. The fungal transformation expression vector, pBARGPE1 was provided by Dr. Krishna Podila, Michigan Technological University. The vector had the Aspergillus nidulans gpdA promoter and the A. nidulans trpC terminator flanking a multiple cloning site. The ß-glucuronidase gene (GUS) was provided by Dr. Randy Allen, Texas Tech University. The ...

The transgenic tobacco plants have been generated that express the E. coli beta-glucuronidase (GUS) gene under control of the promoter from the rice proliferating cell nuclear antigen (PCNA, DNA polymerase auxiliary protein) gene. GUS expression detected in situ by staining with the chromogenic substrate, 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide (X-Gluc), was restricted to meristems in the...

'Mexican' lime (Citrus aurantifolia Swingle) was transformed with constructs that contained chimeric promoter-gus gene fusions of phloem-specific rolC promoter of Agrobacterium rhizogenes, Arabidopsis thaliana sucrose-H(+) symporter (AtSUC2) gene promoter of Arabidopsis thaliana, rice tungro bacilliform virus (RTBV) promoter and sucrose synthase l (RSs1) gene promoter of Oryza sativa (rice). Hi...

The expression and secretion of acid phosphatase (APase) was investigated in Indian mustard (Brassica juncea L. Czern.) plants using sensitive in vitro and activity gel assays. Phosphorus (P) starvation induced two APases in Indian mustard roots, only one of which was secreted. Northern-blot analysis indicated transcriptional regulation of APase expression. Polymerase chain reaction and Souther...

The characterization of organ/tissue-specific promoters is of great interest to transgenic production. The construction of expression cassettes containing tissue-specific promoters is a viable alternative to limited transgene expression to specific organs and cell types. In this context, the purpose of this study was to functionally characterize the promoter of a Eucalyptus grandis gene encodin...