Drought tolerance induced by sound in Arabidopsis plants.

We examined the responses of sound-treated arabidopsis adult plants to water deprivation and the associated changes on gene expression. The survival of drought-induced plants was significantly higher in the sound treated plants (24,8%) compared with plants kept in silence (13,3%). RNA-seq revealed significant upregulation of 87 genes including 32 genes involved in abiotic stress responses, 31 involved in pathogen responses, 11 involved in oxidation-reduction processes, 5 involved in the regulation of transcription, 2 genes involved in protein phosphorylation/dephosphorylation and 13 involved in jasmonic acid or ethylene synthesis or responses. In addition, 2 genes involved in the responses to mechanical stimulus were also induced by sound, suggesting that touch and sound have at least partially common perception and signaling events.

Use of ultrasonication to increase germination rates of Arabidopsis seeds.

BACKGROUND: Arabidopsis thaliana is widely used as model organism in plant biology. Although not of agronomic significance, it offers important advantages for basic research in genetics and molecular biology including the availability of a large number of mutants and genetically modified lines. However, Arabidopsis seed longevity is limited and seeds stored for more than 10 years usually show a very low capacity for germination. RESULTS: The influence of ultrasonic stimulation was investigated on the germination of A. thaliana L. seeds. All experiments have been performed using a frequency of 45 kHz at constant temperature (24 °C). No germination rate differences were observed when using freshly collected seeds. However, using artificially deteriorated seeds, our results show that short ultrasonic stimulation (<1 min) significantly increased germination. Ultrasonic stimulation application of 30 s is the optimal treatment. A significant increase in the germination rate was also verified in naturally aged seeds after ultrasonic stimulation. Scanning electron microscopy observations showed an increase in the presence of pores in the seed coat after sonication that may be the cause, at least in part, of the increase in germination. The ultrasound treated seeds developed normally to mature fertile plants. CONCLUSIONS: Ultrasound technology can be used to enhance the germination process of old Arabidopsis seeds without negatively affecting seedling development. This effect seems to be, at least in part, due to the opening of pores in the seed coat. The use of ultrasonic stimulation in Arabidopsis seeds may contribute to the recovering of long time stored lines.

Genetic diversity of maize germplasm assessed by retrotransposon-based markers.

Maize is one of the most important crops and also a model for grass genome research. Transposable elements comprise over 78% of the maize genome and their ability to generate new copies makes them good potential markers. Interretrotransposon-amplified polymorphism (IRAP) and retrotransposon microsatellite amplified polymorphism (REMAP) protocols were used for the first time in maize to study the genetic variability between maize cultivars. Ten PCR primers were selected based on a systematic analysis of the sequence conservation in the extremities of different high copy number transposable elements, whereas one primer was chosen based on a microsatellite sequence. Of the 16 primer combinations tested, 14 produced polymorphic bands. These markers were used to identify genetic similarity among 20 maize cultivars selected by their different kernel oil content. Genetic similarity analysis was performed based on the polymorphic band profiles and dendrograms were developed by the unweighted pair-group method with arithmetic averages. Clustering technique revealed that samples were grouped into three clusters that differed in their kernel oil content and size, and in their relative embryo size. In the current investigation, there is evidence that IRAP/REMAP may be useful as markers in maize.

The evolutionary conserved oil body associated protein OBAP1 participates in the regulation of oil body size.

A transcriptomic approach has been used to identify genes predominantly expressed in maize (Zea mays) scutellum during maturation. One of the identified genes is oil body associated protein1 (obap1), which is transcribed during seed maturation predominantly in the scutellum, and its expression decreases rapidly after germination. Proteins similar to OBAP1 are present in all plants, including primitive plants and mosses, and in some fungi and bacteria. In plants, obap genes are divided in two subfamilies. Arabidopsis (Arabidopsis thaliana) genome contains five genes coding for OBAP proteins. Arabidopsis OBAP1a protein is accumulated during seed maturation and disappears after germination. Agroinfiltration of tobacco (Nicotiana benthamiana) epidermal leaf cells with fusions of OBAP1 to yellow fluorescent protein and immunogold labeling of embryo transmission electron microscopy sections showed that OBAP1 protein is mainly localized in the surface of the oil bodies. OBAP1 protein was detected in the oil body cellular fraction of Arabidopsis embryos. Deletion analyses demonstrate that the most hydrophilic part of the protein is responsible for the oil body localization, which suggests an indirect interaction of OBAP1 with other proteins in the oil body surface. An Arabidopsis mutant with a transfer DNA inserted in the second exon of the obap1a gene and an RNA interference line against the same gene showed a decrease in the germination rate, a decrease in seed oil content, and changes in fatty acid composition, and their embryos have few, big, and irregular oil bodies compared with the wild type. Taken together, our findings suggest that OBAP1 protein is involved in the stability of oil bodies.

ZmPTR1, a maize peptide transporter expressed in the epithelial cells of the scutellum during germination.

In plants, peptide transporter/nitrate transporter 1 (PTR/NRT1) family proteins transport a variety of substrates such as nitrate, di- and tripepetides, auxin and carboxylates across membranes. We isolated and characterized ZmPTR1, a maize member of this family. ZmPTR1 protein sequence is highly homologous to the previously characterized di- and tripeptide Arabidopsis transporters AtPTR2, AtPTR4 and AtPTR6. ZmPTR1 gene is expressed in the cells of the scutellar epithelium during germination and, to a less extent, in the radicle and the hypocotyl. Arabidopsis thaliana lines overexpressing ZmPTR1 performed better than control plants when grown on a medium with Ala-Ala dipeptide as the unique N source. Our results suggest that ZmPTR1 plays a role in the transport into the embryo of the small peptides produced during enzymatic hydrolysis of the storage proteins in the endosperm.

Expression profile of maize (Zea mays) scutellar epithelium during imbibition.

The scutellum is a shield-shaped structure surrounding the embryo axis in grass species. The scutellar epithelium (Sep) is a monolayer of cells in contact with the endosperm. The Sep plays an important role during seed germination in the secretion of gibberellins and hydrolytic enzymes and in the transport of the hydrolized products to the growing embryo. We identified 30 genes predominantly expressed after imbibition in the Sep as compared to other parts of the scutellum. A high proportion of these genes is involved in metabolic processes. Some other identified genes are involved in the synthesis or modification of cell walls, which may be reflected in the changes of cell shape and cell wall composition that can be observed during imbibition. One of the genes encodes a proteinase that belongs to a proteinase family typical of carnivorous plants. Almost nothing is known about their role in other plants or organs, but the scutellar presence may point to a "digestive" function during germination. Genes involved in the production of energy and the transport of peptides were also identified.

Genetic, molecular and cellular approaches to the analysis of maize embryo development.

The development of embryo structures in plants is essential for the formation of the adult plant organs. In cereals, this process has distinct features which have attracted attention from different points of view. Differential gene expression analyses have been used in order to identify genes useful as molecular markers of certain physiological, molecular or developmental processes. Several maize mutants affected in embryo development have been isolated, but only a fraction of them have been characterized at the molecular level. Molecular markers can be useful in the characterization of embryo defective mutants. Here, we describe the different techniques used in the identification of molecular marker genes for embryo development. We describe in more detail some groups of genes coding for cell wall proteins. We also describe the application of these molecular markers in the characterization of some embryo mutants.