Methylglyoxal induces stress signaling and promotes the germination of maize at low temperature.

Maize is sensitive to cold injury, especially during germination. Since cold causes oxidative stress, compounds that promote the accumulation of free radical forms, such as the reactive aldehyde (RA) methylglyoxal (MG), may be suitable to trigger a systemic defense response. In this study, maize seeds were soaked in MG solution for one night at room temperature, before germination test at 13°C. The exogenous MG enhanced the germination and photosynthetic performance of maize at low temperature. Transcriptome analysis, hormonal, and flavonoid profiling indicated MG-induced changes in photosystem antenna proteins, pigments, late embryogenesis abundant proteins, abscisic acid (ABA) derivatives, chaperons, and certain dihydroflavonols, members of the phenylpropanoid pathway. MG-response of the two maize cultivars (A654 and Cm174) were somewhat different, but we recorded higher endogenous hydrogen peroxide (H2 O2 ) and lower nitric oxide (NO) level in at least one of the treated genotypes. These secondary signal molecules may provoke some of the changes in the hormonal, metabolic and gene expression profile. Decreased auxin transport, but increased ABA degradation and cytokinin and jasmonic acid (JA) synthesis, as well as an altered carbohydrate metabolism and transport (amylases, invertases, and SWEET transporters) could have promoted germination of MG-pretreated seeds. While LEA accumulation could have protected against osmotic stress and catalase expression and production of many antioxidants, like para-hydroxybenzoic acid (p-HBA) and anthocyanins may have balanced the oxidative environment for maize germination. Our results showed that MG-pretreatment could be an effective way to promote cold germination and its effect was more pronounced in the originally cold-sensitive maize genotype.

Global transcriptome and targeted metabolite analyses of roots reveal different defence mechanisms against Ralstonia solanacearum infection in two resistant potato cultivars.

Ralstonia solanacearum (Rs), the causal agent of bacterial wilt disease in an unusually wide range of host plants, including potato (Solanum tuberosum), is one of the most destructive phytopathogens that seriously reduces crop yields worldwide. Identification of defence mechanisms underlying bacterial wilt resistance is a prerequisite for biotechnological approaches to resistance breeding. Resistance to Rs has been reported only in a few potato landraces and cultivars. Our in vitro inoculation bioassays confirmed that the cultivars 'Calalo Gaspar' (CG) and 'Cruza 148' (CR) are resistant to Rs infection. Comparative transcriptome analyses of CG and CR roots, as well as of the roots of an Rs-susceptible cultivar, 'Désirée' (DES), were carried out two days after Rs infection, in parallel with their respective noninfected controls. In CR and DES, the upregulation of chitin interactions and cell wall-related genes was detected. The phenylpropanoid biosynthesis and glutathione metabolism pathways were induced only in CR, as confirmed by high levels of lignification over the whole stele in CR roots six days after Rs infection. At the same time, Rs infection greatly increased the concentrations of chlorogenic acid and quercetin derivatives in CG roots as it was detected using ultra-performance liquid chromatography - tandem mass spectrometry. Characteristic increases in the expression of MAP kinase signalling pathway genes and in the concentrations of jasmonic, salicylic, abscisic and indoleacetic acid were measured in DES roots. These results indicate different Rs defence mechanisms in the two resistant potato cultivars and a different response to Rs infection in the susceptible cultivar.

Exogenous methylglyoxal enhances the reactive aldehyde detoxification capability and frost-hardiness of wheat.

Cold-acclimation is essential for the development of adequate frost-hardiness in cereals and therefore sudden freezes can cause considerable damage to the canopy. However, timely adding of an appropriate signal in the absence of cold acclimation may also harden wheat for the upcoming freeze. The feasibility of the promising signal molecule methylglyoxal was tested here for such applications and the signal mechanism was studied in bread wheat (Triticum aestivum L.) and durum wheat (Triticum turgidum L. ssp. durum). Spraying with 10 mM methylglyoxal did not decrease the fresh weight and photosynthetic parameters in most wheat varieties at growth temperature (21 °C). Photosynthetic parameters even improved and chlorophyll content increased in some cases. Increased transcript level of glutathione-S-transferases and omega-3 fatty acid desaturases was detected by qPCR 6 h after the last methylglyoxal spray. Aldo-keto reductase and glyoxalase enzyme activities, as well as sorbitol content of wheat plants increased 24 h after the last 10 mM methylglyoxal spray in most of the cultivars. These mechanisms may explain the increased freezing survival of methylglyoxal pretreated wheat plants from less than 10% to over 30%. Our results demonstrate that exogenous methylglyoxal treatment can be safely added to wheat plants as preparatory treatment without detrimental effects but inducing some of the stress-protective mechanisms, which contribute to frost-hardiness.

Current and possible approaches for improving photosynthetic efficiency.

One of the most important tasks laying ahead today's biotechnology is to improve crop productivity with the aim of meeting increased food and energy demands of humankind. Plant productivity depends on many genetic factors, including life cycle, harvest index, stress tolerance and photosynthetic activity. Many approaches were already tested or suggested to improve either. Limitations of photosynthesis have also been uncovered and efforts been taken to increase its efficiency. Examples include decreasing photosynthetic antennae size, increasing the photosynthetically available light spectrum, countering oxygenase activity of Rubisco by implementing C4 photosynthesis to C3 plants and altering source to sink transport of metabolites. A natural and effective photosynthetic adaptation, the sugar alcohol metabolism got however remarkably little attention in the last years, despite being comparably efficient as C4, and can be considered easier to introduce to new species. We also propose root to shoot carbon-dioxide transport as a means to improve photosynthetic performance and drought tolerance at the same time. Different suggestions and successful examples are covered here for improving plant photosynthesis as well as novel perspectives are presented for future research.

Cold inducible promoter driven Cre-lox system proved to be highly efficient for marker gene excision in transgenic barley.

A Cre-lox based auto-excision strategy has been adapted for barley, capable of cre and selectable marker gene (SMG) removal. The cold inducible wheat promoter called wcs120 was utilised for driving Cre expression. The binary vector was carrying the transgene (uidA) and a so called 'recombination cassette' flanked by the lox sequences. This part included both the recombinase gene and the SMG (bar) under the control of a constitutive promoter. T0, T1 and T2 transgenic plants were subjected to low temperature (at 4°C, 10°C and 12°C) at different developmental stages to induce recombination. The presence of uidA, cre, and bar genes and recombination footprints were studied by PCR and DNA sequencing, while cre transcription was followed by qRT-PCR. These analyses indicated that, cold treatment of the germinating seeds (4°C for 3days) followed by plant growing at higher temperature (24°C) has been the most efficient (90-100%), and this treatment lead to heritable changes in the genome. Thermal separation of Cre accumulation (at low temperature) from Cre enzyme activity (at higher temperature) could have prevented the premature excision of its own encoding gene, and lead to high expression level thereby increasing recombination frequency.

Multiple elements controlling the expression of wheat high molecular weight glutenin paralogs.

Analysis of gene expression data generated by high-throughput microarray transcript profiling experiments coupled with cis-regulatory elements enrichment study and cluster analysis can be used to define modular gene programs and regulatory networks. Unfortunately, the high molecular weight glutenin subunits of wheat (Triticum aestivum) are more similar than microarray data alone would allow to distinguish between the three homoeologous gene pairs. However, combining complementary DNA (cDNA) expression libraries with microarray data, a co-expressional network was built that highlighted the hidden differences between these highly similar genes. Duplex clusters of cis-regulatory elements were used to focus the co-expressional network of transcription factors to the putative regulatory network of Glu-1 genes. The focused network helped to identify several transcriptional gene programs in the endosperm. Many of these programs demonstrated a conserved temporal pattern across the studied genotypes; however, few others showed variance. Based on this network, transient gene expression assays were performed with mutated promoters to inspect the control of tissue specificity. Results indicated that the interactions of the ABRE│CBF cluster with distal promoter regions may have a dual role in regulation by both recruiting the transcription complex as well as suppressing it in non-endosperm tissue. A putative model of regulation is discussed.