Mercury contamination in food can pose serious health risks to consumers and coal-fired power plants have been identified as the major source of mercury emissions. To assess the current state of mercury pollution in food crops grown near coal-fired power plants, we measured the total mercury concentration in vegetables and grain crops collected from farms located near two coal-fired power plants. We found that 79% of vegetable samples and 67% of grain samples exceeded the PTWI’s food safety standards. The mercury concentrations of soil samples were negatively correlated with distances from the studied coal-fired power plants, and the mercury contents in lettuce, amaranth, water spinach, cowpea and rice samples were correlated with the mercury contents in soil samples, respectively. Also, the mercury concentrations in vegetable leaves were much higher than those in roots and the mercury content of vegetable leaves decreased significantly after water rinses. Our calculation suggests that probable weekly intake of mercury for local residents, assuming all of their vegetables and grains are from their own farmland, may exceed the toxicologically tolerable values allowed, and therefore long-term consumptions of these contaminated vegetables and grains may pose serious health risks.
Developing CRISPR/Cas9-mediated non-transgenic mutants in asexually propagated perennial crop plants is challenging but highly desirable. Here, we report a highly useful method using an Agrobacterium-mediated transient CRISPR/Cas9 gene expression system to create non-transgenic mutant plants without the need for sexual segregation. We have also developed a rapid, cost-effective, and high-throughput mutant screening protocol based on Illumina sequencing followed by high-resolution melting (HRM) analysis. Using tetraploid tobacco as a model species and the phytoene desaturase (PDS) gene as a target, we successfully created and expediently identified mutant plants, which were verified as tetra-allelic mutants. We produced pds mutant shoots at a rate of 47.5% from tobacco leaf explants, without the use of antibiotic selection. Among these pds plants, 17.2% were confirmed to be non-transgenic, for an overall non-transgenic mutation rate of 8.2%. Our method is reliable and effective in creating non-transgenic mutant plants without the need to segregate out transgenes through sexual reproduction. This method should be applicable to many economically important, heterozygous, perennial crop species that are more difficult to regenerate.
Summary: Pollen‐ and seed‐mediated transgene flow is a concern in plant biotechnology. We report here a highly efficient ‘genetically modified (GM)‐gene‐deletor’ system to remove all functional transgenes from pollen, seed or both. With the three pollen‐ and/or seed‐specific gene promoters tested, the phage CRE/loxP or yeast FLP/FRT system alone was inefficient in excising transgenes from tobacco pollen and/or seed, with no transgenic event having 100% efficiency. When loxP‐FRT fusion sequences were used as recognition sites, simultaneous expression of both FLP and CRE reduced the average excision efficiency, but the expression of FLP or CRE alone increased the average excision efficiency, with many transgenic events being 100% efficient based on more than 25 000 T1 progeny examined per event. The ‘GM‐gene‐deletor’ reported here may be used to produce ‘non‐transgenic’ pollen and/or seed from transgenic plants and to provide a bioconfinement tool for transgenic crops and perennials, with special applicability towards vegetatively propagated plants and trees.
Horticulture Research is a new Open Access, fully peer-reviewed journal which will publish the best and most interesting research focusing on all major horticultural crops from around the world.
Gibberellins (GAs) play a crucial role in growth and development of the tomato fruit. Previously published studies focusing on the effect of GAs on tomato fruits used chemical treatments, constitutive overexpression or silencing of GA biosynthetic and catabolic genes globally throughout the plant. Fruit-specific overexpression of GA catabolic enzyme genes GA2-oxidases (GA2oxs), however, may provide an alternative method to study the role of endogenous GAs on the fruit development. In this study, we have identified 11 SlGA2ox proteins in tomato that are classified into three subgroups. Motif analysis and multiple sequence alignments have demonstrated that all SlGA2oxs, except SlGA2ox10, have similar motif compositions and high-sequence conservation. Quantitative reverse transcription-PCR analysis has showed that SlGA2oxs exhibit differential expression patterns in tomato fruits at different developmental stages. When the fruit-specific promoter TFM7 was used to control the expression of SlGA2ox1, we observed no changes in growth and development of vegetative organs. However, fruit weight, seed number and germination rate were significantly affected. We also treated tomato fruits with GA biosynthesis inhibitor and observed phenotypes similar to those of the transgenic fruits. Furthermore, we have demonstrated that expression of cell expansion and GA responsive genes were downregulated in transgenic tomato fruits, supporting that overexpression of the SlGA2ox1 leads to reduction in endogenous GAs. This study provides additional evidence that endogenous GAs and the SlGA2ox1 gene play an important role in controlling on fruit weight, seed development and germination in tomato plant.