Microalgae are considered one of the most promising feedstocks for biofuels. The productivity of these photosynthetic microorganisms in converting carbon dioxide into carbon-rich lipids, only a step or two away from biodiesel, greatly exceeds that of agricultural oleaginous crops, without competing for arable land. Worldwide, research and demonstration programs are being carried out to develop the technology needed to expand algal lipid production from a craft to a major industrial process. Although microalgae are not yet produced at large scale for bulk applications, recent advances—particularly in the methods of systems biology, genetic engineering, and biorefining—present opportunities to develop this process in a sustainable and economical way within the next 10 to 15 years.

https://science.sciencemag.org/content/sci/329/5993/796.full.pdf

An outlook on microalgal biofuels.

▪ Abstract Plants respond to herbivore attack with a bewildering array of responses, broadly categorized as direct and indirect defenses, and tolerance. Plant-herbivore interactions are played out on spatial scales that include the cellular responses, well-studied in plant-pathogen interactions, as well as responses that function at whole-plant and community levels. The plant's wound response plays a central role but is frequently altered by insect-specific elicitors, giving plants the potential to optimize their defenses. In this review, we emphasize studies that advance the molecular understanding of elicited direct and indirect defenses and include verifications with insect bioassays. Large-scale transcriptional changes accompany insect-induced resistance, which is organized into specific temporal and spatial patterns and points to the existence of herbivore-specific trans-activating elements orchestrating the responses. Such organizational elements could help elucidate the molecular control over the d...

Plant responses to insect herbivory: The emerging molecular analysis

Phenolic secondary metabolites play an important role in plant-derived food quality, as they affect quality characteristics such as appearance, flavour and health-promoting properties. Their content in foods is affected by many factors that influence phenolic stability, biosynthesis and degradation. In terms of their biosynthesis the key enzyme phenylalanine ammonia-lyase (PAL) is especially relevant, as it can be induced by different stress (environmental) conditions. In addition, polyphenol oxidases (PPO) and peroxidases (POD) are the main enzymes responsible for quality loss due to phenolic degradation. The different factors affecting phenolic-related food quality are reviewed. These include internal (genetic) and environmental (agronomic) factors, technological treatments applied during postharvest storage of fruits and vegetables, as well as processing and storage of the processed products. The different strategies that are required to either maintain or enhance the phenolic-related quality of foods are critically reviewed. Genetic modification designed to decrease polyphenol oxidases or peroxidases is not always a feasible method, owing to side problems related to the growth and defence of the plant. Agronomic treatments can be used to enhance the phenolic content and pigmentation of fruits and vegetables, although the information available on this topic is very scarce and even contradictory. Some postharvest treatments (cold storage, controlled or modified atmospheres, etc) can also improve phenolic-related quality, as well as new processing methods such as irradiation (gamma, UV), high-field electric pulses, high hydrostatic pressures and microwaves. Les composes phenoliques, metabolites secondaires des plantes, jouent un role dans la qualite des fruits, au niveau de l'apparence, de la flaveur et des qualites nutritives. L'enzyme cle de leur biosynthese est la phenylalanine ammonium-lyase, regulee par les facteurs environnementaux. La polyphenol oxydase et la peroxydase sont responsables des degradations donc des pertes de qualite. Les differents facteurs internes (genetiques) et externes (agronomiques) affectant la qualite des produits vegetaux sont etudies. Les differentes mesures envisageables (modification genetique, traitements chimiques ou physiques apres recolte) pour modifier le contenu en compose phenoliques et donc la qualite des produits vegetaux sont etudies.

Phenolic compounds and related enzymes as determinants of quality in fruits and vegetables

There are currently intensive global research efforts aimed at increasing and modifying the accumulation of lipids, alcohols, hydrocarbons, polysaccharides, and other energy storage compounds in photosynthetic organisms, yeast, and bacteria through genetic engineering. Many improvements have been realized, including increased lipid and carbohydrate production, improved H2 yields, and the diversion of central metabolic intermediates into fungible biofuels. Photosynthetic microorganisms are attracting considerable interest within these efforts due to their relatively high photosynthetic conversion efficiencies, diverse metabolic capabilities, superior growth rates, and ability to store or secrete energy-rich hydrocarbons. Relative to cyanobacteria, eukaryotic microalgae possess several unique metabolic attributes of relevance to biofuel production, including the accumulation of significant quantities of triacylglycerol; the synthesis of storage starch (amylopectin and amylose), which is similar to that found in higher plants; and the ability to efficiently couple photosynthetic electron transport to H2 production. Although the application of genetic engineering to improve energy production phenotypes in eukaryotic microalgae is in its infancy, significant advances in the development of genetic manipulation tools have recently been achieved with microalgal model systems and are being used to manipulate central carbon metabolism in these organisms. It is likely that many of these advances can be extended to industrially relevant organisms. This review is focused on potential avenues of genetic engineering that may be undertaken in order to improve microalgae as a biofuel platform for the production of biohydrogen, starch-derived alcohols, diesel fuel surrogates, and/or alkanes.

Genetic Engineering of Algae for Enhanced Biofuel Production

An analysis of changes in global gene expression patterns during developmental leaf senescence in Arabidopsis has identified more than 800 genes that show a reproducible increase in transcript abundance. This extensive change illustrates the dramatic alterations in cell metabolism that underpin the developmental transition from a photosynthetically active leaf to a senescing organ which functions as a source of mobilizable nutrients. Comparison of changes in gene expression patterns during natural leaf senescence with those identified, when senescence is artificially induced in leaves induced to senesce by darkness or during sucrose starvation-induced senescence in cell suspension cultures, has shown not only similarities but also considerable differences. The data suggest that alternative pathways for essential metabolic processes such as nitrogen mobilization are used in different senescent systems. Gene expression patterns in the senescent cell suspension cultures are more similar to those for dark-induced senescence and this may be a consequence of sugar starvation in both tissues. Gene expression analysis in senescing leaves of plant lines defective in signalling pathways involving salicylic acid (SA), jasmonic acid (JA) and ethylene has shown that these three pathways are all required for expression of many genes during developmental senescence. The JA/ethylene pathways also appear to operate in regulating gene expression in dark-induced and cell suspension senescence whereas the SA pathway is not involved. The importance of the SA pathway in the senescence process is illustrated by the discovery that developmental leaf senescence, but not dark-induced senescence, is delayed in plants defective in the SA pathway.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-313X.2005.02399.x

Comparative transcriptome analysis reveals significant differences in gene expression and signalling pathways between developmental and dark/starvation-induced senescence in Arabidopsis.

We report the recombinant preparation from Escherichia coli cells of samples of two closely related, small, secreted cysteine-rich plant peptides: rapid alkalinization factor 1 (RALF1) and rapid alkalinization factor 8 (RALF8). Purified samples of the native sequence of RALF8 exhibited well-resolved nuclear magnetic resonance (NMR) spectra and also biological activity through interaction with a plant receptor kinase, cytoplasmic calcium mobilization, and in vivo root growth suppression. By contrast, RALF1 could only be isolated from inclusion bodies as a construct containing an N-terminal His-tag; its poorly resolved NMR spectrum was indicative of aggregation. We prepared samples of the RALF8 peptide labeled with 15 N and 13 C for NMR analysis and obtained near complete 1 H, 13 C, and 15 N NMR assignments; determined the disulfide pairing of its four cysteine residues; and examined its solution structure. RALF8 is mostly disordered except for the two loops spanned by each of its two disulfide bridges.

https://www.onlinelibrary.wiley.com/doi/pdf/10.1002/pro.3628

Function and solution structure of the Arabidopsis thaliana RALF8 peptide.

In Arabidopsis, defense signaling is triggered by the perception of conserved molecular patterns by pattern recognition receptors (PRRs). Signal transduction from the PRRs requires members of a family of Receptor-Like Cytoplasmic Kinases (RLCKs). Previously, we described one such RLCK, PTI Compromised Receptor-Like Cytoplasmic Kinase 1 (PCRK1) that is important for immunity induced by Microbe Associated Molecular Patterns (MAMPs) as well as Damage Associated Molecular Patterns (DAMPs). In this study, we measured the growth of Pma ES4326 in double mutants carrying pcrk1 together with the salicylic acid (SA) biosynthesis mutation sid2–2 or the jasmonic acid (JA) receptor mutation coi1–1, showing that the function of PCRK1 is SA independent but may be partially dependent on JA. Mutation of phosphorylated serine residues S232, S233 and S237 compromised the immune signaling function of PCRK1.

Miyoshi Haruta

Papers

Function and solution structure of the Arabidopsis thaliana RALF8 peptide.

Functional characterization of PCRK1, a putative protein kinase with a role in immunity

Effect of Transformation by the Agrobacterium Method on the Proteins and Secondary Metabolites of Apple and Tobacco Shoots