Use of systemic biofertilizers in sugarcane results in highly reproducible increments in yield and quality of harvests.

The utilization of a novel (systemic) biofertilizer containing Pseudomonas fluorescens, Azospirillum brasilense, and Bacillus subtilis and possessing the technology to facilitate the entry of bacteria through the stomata, was evaluated at three localities in Mexico (Potrero Nuevo, Veracruz; Ameca, Jalisco; and Champotón, Campeche) in two sugarcane varieties (NCO-310 and Mex 57-473) at different time scales. Inoculation of the systemic biofertilizer was imposed over the local agricultural management of the sugarcane; chemical fertilization of the experimental parcels at Potrero Nuevo was done using 70-20-20 and 120-80-80 at Ameca and Champotón. Three doses of the biofertilizer per hectare were applied during the annual productive cycle of sugarcane at each site; one year at Potrero Nuevo and Champotón; and six years at Ameca. The annual sugarcane yield was evaluated at each site. Additionally, sugar quality (°Brix or sucrose content) was evaluated at the three localities, while different variables of stalk performance were also measured at Ameca and Champotón. Our data provide evidence that this systemic biofertilizer consistently and reliably increased the sugarcane yield at all localities during the time of evaluation, ranging from 73.7 tons ha-1 at Potrero Nuevo (2.5 times increase; P < 0.05) and 77.7 tons ha-1 at Ameca (1.9 times increase; P < 0.05) to 23.8 tons ha-1 at Champotón (1.4 times increase; P < 0.05). This increase in sugarcane biomass was related to increased tillering rather than increased stalk height or diameter. This novel biological product improved the sugarcane quality in terms of °Brix (P < 0.05, 2.6° difference) and sucrose content (P < 0.5, 0.7% difference).

Identification of miRNA from Bouteloua gracilis, a drought tolerant grass, by deep sequencing and their in silico analysis.

BACKGROUND: MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate signal transduction, development, metabolism, and stress responses in plants through post-transcriptional degradation and/or translational repression of target mRNAs. Several studies have addressed the role of miRNAs in model plant species, but miRNA expression and function in economically important forage crops, such as Bouteloua gracilis (Poaceae), a high-quality and drought-resistant grass distributed in semiarid regions of the United States and northern Mexico remain unknown. RESULTS: We applied high-throughput sequencing technology and bioinformatics analysis and identified 31 conserved miRNA families and 53 novel putative miRNAs with different abundance of reads in chlorophyllic cell cultures derived from B. gracilis. Some conserved miRNA families were highly abundant and possessed predicted targets involved in metabolism, plant growth and development, and stress responses. We also predicted additional identified novel miRNAs with specific targets, including B. gracilis ESTs, which were detected under drought stress conditions. CONCLUSIONS: Here we report 31 conserved miRNA families and 53 putative novel miRNAs in B. gracilis. Our results suggested the presence of regulatory miRNAs involved in modulating physiological and stress responses in this grass species.

A reliable method for spectrophotometric determination of glycine betaine in cell suspension and other systems.

Glycine betaine is a quaternary ammonium compound that accumulates in a large variety of species in response to different types of stress. Glycine betaine counteracts adverse effects caused by abiotic factors, preventing the denaturation and inactivation of proteins. Thus, its determination is important, particularly for scientists focused on relating structural, biochemical, physiological, and/or molecular responses to plant water status. In the current work, we optimized the periodide technique for the determination of glycine betaine levels. This modification permitted large numbers of samples taken from a chlorophyllic cell line of the grass Bouteloua gracilis to be analyzed. Growth kinetics were assessed using the chlorophyllic suspension to determine glycine betaine levels in control (no stress) cells and cells osmotically stressed with 14 or 21% polyethylene glycol 8000. After glycine extraction, different wavelengths and reading times were evaluated in a spectrophotometer to determine the optimal quantification conditions for this osmolyte. Optimal results were obtained when readings were taken at a wavelength of 290 nm at 48 h after dissolving glycine betaine crystals in dichloroethane. We expect this modification to provide a simple, rapid, reliable, and cheap method for glycine betaine determination in plant samples and cell suspension cultures.

Chlorophyll accumulation is enhanced by osmotic stress in graminaceous chlorophyllic cells.

We have developed a new chlorophyllic cell line ('TADH-XO') from the highly water stress tolerant grass Bouteloua gracilis (blue grama). When grown under normal (non-stress) conditions, this new cell line accumulates higher levels of chlorophyll (up to 368.1 microg total chlorophyll g(-1) FW) than a previously obtained cell line ('TIANSJ98'). Both cell lines are capable of developing substantially higher amounts of chlorophyll when subjected to osmotic stress. In order to explain these changes in the chlorophyll kinetics of the chlorophyllic cells, we analyzed the following population variables in cells subjected to polyethylene glycol 8000-induced osmotic stress: growth, viability, chlorophyll (total, 'a' and 'b'), cell size, percentage of green cells and chloroplast (number and size). Although previous studies in some chlorophyllic cells of dicots have already reported that chlorophyll increases under saline stress, in this report we show that, at least in this graminaceous cell line, the increase in chlorophyll is an immediate and proportional response to the osmotic stress applied and not the result of a progressive adaptation process. Consistent with previous studies, the increase in chlorophyll accumulation could be the result of chloroplast development (increased thylakoid number per chloroplast). On the basis of our results, the increases in chlorophyll accumulation previously observed in salt-adapted dicot cells may be the result of the osmotic shock (water deficit), rather than the ionic effect of salt on the physiology of chlorophyllic cells of dicots. Under the cell population experimental approach we followed, our study provides important insights related to the physiological behavior of chlorophyllic cells subjected to osmotic stress.