Background: The elucidation of gene expression patterns leads to a better understanding of biological processes. Real-time quantitative RT-PCR has become the standard method for in-depth studies of gene expression. A biologically meaningful reporting of target mRNA quantities requires accurate and reliable normalization in order to identify real gene-specific variation. The purpose of normalization is to control several variables such as different amounts and quality of starting material, variable enzymatic efficiencies of retrotranscription from RNA to cDNA, or differences between tissues or cells in overall transcriptional activity. The validity of a housekeeping gene as endogenous control relies on the stability of its expression level across the sample panel being analysed. In the present report we describe the first systematic evaluation of potential internal controls during tomato development process to identify which are the most reliable for transcript quantification by real-time RT-PCR.

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Biostimulants (BSs) are probably one of the most promising alternatives nowadays to cope with yield losses caused by plant stress, which are intensified by climate change. Biostimulants comprise many different compounds with positive effects on plants, excluding pesticides and chemical fertilisers. Usually mixtures such as lixiviates from proteins or algal extracts have been used, but currently companies are interested in more specific compounds that are capable of increasing tolerance against abiotic stress. Individual application of a pure active compound offers researchers the opportunity to better standarise formulations, learn more about the plant defence process itself and assist the agrochemical industry in the development of new products. This review attempts to summarise the state of the art regarding various families of organic compounds and their mode/mechanism of action as BSs, and how they can help maximise agricultural yields under stress conditions aggravated by climate change.

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Water scarcity is one of the main challenges faced by modern agriculture, leading to a substantial drop in crop productivity and a threat to food security. Thus, novel agricultural approaches are necessary and urgent to face this problem. Some natural compounds such as amino acids have been shown to increase yield and mitigate the effects of drought stress. In this study, we demonstrate that the application of pyroglutamic acid (PG) is capable of increasing lettuce yield, under field conditions with 30% less than optimal irrigation. PG treatment showed a clearly protective effect in stressed plants, enhancing their fresh weight by 37% and yield by 31%, in comparison to untreated plants. PG appears to promote drought tolerance effects in deficit irrigated lettuce plants, with several advantages. It acts by enhancing photosynthesis rate and antioxidant defences, while maintaining osmotic and water balance, without toxicity to soil microorganisms. This illustrates the potential use of PG to combat productivity losses due to water scarcity.

SummaryMenadione sodium bisulphite (MSB) is a water-soluble derivative of vitamin K3, or menadione, and has been previously demonstrated to function as a plant defence activator against several pathogens in several plant species. However, there are no reports of the role of this vitamin in the induction of resistance in the plant model Arabidopsis thaliana. In the current study, we demonstrate that MSB induces resistance by priming in Arabidopsis against the virulent strain Pseudomonas syringae pv.tomato DC3000 (Pto) without inducing necrosis or visible damage. Changes in gene expression in response to 0.2 mM MSB were analysed in Arabidopsis at 3, 6 and 24 h post-treatment using microarray technology. In general, the treatment with MSB does not correlate with other publicly available data, thus MSB produces a unique molecular footprint. We observed 158 differentially regulated genes among all the possible trends. More up-regulated genes are included in categories such as 'response to stress' than the background, and the behaviour of these genes in different treatments confirms their role in response to biotic and abiotic stress. In addition, there is an over-representation of the G-box in their promoters. Some interesting functions are represented among the individual up-regulated genes, such as glutathione S-transferases, transcription factors (including putative regulators of the G-box) and cytochrome P450s. This work provides a wide insight into the molecular cues underlying the effect of MSB as a plant resistance inducer.

Pretreatment of the first true leaves of oilseed rape plants ( Brassica napus cv. Bristol) with menadione sodium bisulphite (MSB) locally and systemically induced resistance, as shown by reduced lesion size and number, to infection by the fungal pathogen Leptosphaeria maculans , the causal agent of stem canker. Using a known systemic activator of salicylic aciddependent PR-1 induction, acibenzolar-S -methyl (BTH; S -methylbenzo[1,2,3]thiadiazole-7-carbothiate) as a comparison, real-time PCR expression analysis of genes encoding a pathogenesis-related protein 1 ( PR-1 ) and an ascorbate peroxidase ( APX ) demonstrated a systemic enhancement of APX expression in MSB-pretreated plants, with no effect on PR-1 expression, suggesting augmented reactive oxygen species production in MSB-pretreated plants. The results demonstrate MSB to be an effective resistance activator in oilseed rape, and potentially useful for the control of stem canker.