QTL were identified for root architectural traits in maize. Root architectural traits, including the number, length, orientation, and branching of the principal root classes, influence plant function by determining the spatial and temporal domains of soil exploration. To characterize phenotypic patterns and their genetic control, three recombinant inbred populations of maize were grown for 28 days in solid media in a greenhouse and evaluated for 21 root architectural traits, including length, number, diameter, and branching of seminal, primary and nodal roots, dry weight of embryonic and nodal systems, and diameter of the nodal root system. Significant phenotypic variation was observed for all traits. Strong correlations were observed among traits in the same root class, particularly for the length of the main root axis and the length of lateral roots. In a principal component analysis, relationships among traits differed slightly for the three families, though vectors grouped together for traits within a given root class, indicating opportunities for more efficient phenotyping. Allometric analysis showed that trajectories of growth for specific traits differ in the three populations. In total, 15 quantitative trait loci (QTL) were identified. QTL are reported for length in multiple root classes, diameter and number of seminal roots, and dry weight of the embryonic and nodal root systems. Phenotypic variation explained by individual QTL ranged from 0.44 % (number of seminal roots, NyH population) to 13.5 % (shoot dry weight, OhW population). Identification of QTL for root architectural traits may be useful for developing genotypes that are better suited to specific soil environments.

QTL mapping and phenotypic variation for root architectural traits in maize (Zea mays L.)

This review covers both methodical aspects and actual applications of ion imaging techniques in plant cell signal research. The methodological section explains the basic principles of fluorescence ion imaging, the impact of modern developments in fluorescence microscopy and introduces the most important fluorescence probes including aequorin and other photoproteins. It critically comments on loading strategies, intracellular compartmentation of probes and calibration procedures. The second part compiles actual research areas where the application of ion imaging procedures has gained substantial achievements and helped to establish new concepts of calcium- and pH-dependent signalling. Examples comprise the hormonal control of stomatal movements, effects of gibberellic and abscisic acids in aleurone cells, elicitation of phytoalexin production, cytosolic pH and cell development, and signatures of Ca2+ as a universal signal in plant cells.

Ion mapping in plant cells--methods and applications in signal transduction research.

Regulated protein secretion is required for malaria parasite life cycle progression and transmission between the mammalian host and mosquito vector. During transmission from the host to the vector, exocytosis of highly specialised secretory vesicles, such as osmiophilic bodies, is key to the dissolution of the red blood cell and parasitophorous vacuole membranes enabling gamete egress. The positioning of adhesins from the TRAP family, from micronemes to the sporozoite surface, is essential for gliding motility of the parasite and transmission from mosquito to mammalian host. Here we identify a conserved role for the putative pantothenate transporter PAT in Plasmodium berghei in vesicle fusion of two distinct classes of vesicles in gametocytes and sporozoites. PAT is a membrane component of osmiophilic bodies in gametocytes and micronemes in sporozoites. Despite normal formation and trafficking of osmiophilic bodies to the cell surface upon activation, PAT-deficient gametes fail to discharge their contents, remain intraerythrocytic and unavailable for fertilisation and further development in the mosquito. Sporozoites lacking PAT fail to secrete TRAP, are immotile and thus unable to infect the subsequent rodent host. Thus, P. berghei PAT appears to regulate exocytosis in two distinct populations of vesicles in two different life cycle forms rather than acting as pantothenic transporter during parasite transmission.

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A Putative Small Solute Transporter Is Responsible for the Secretion of G377 and TRAP-Containing Secretory Vesicles during Plasmodium Gamete Egress and Sporozoite Motility.

Root-root interactions are much more sophisticated than previously thought, yet the mechanisms of belowground neighbor perception remain largely obscure. Genome-wide transcriptome analyses allow detailed insight into plant reactions to environmental cues. A root interaction trial was set up to explore both morphological and whole genome transcriptional responses in roots of Arabidopsis thaliana in the presence or absence of an inferior competitor, Hieracium pilosella. Neighbor perception was indicated by Arabidopsis roots predominantly growing away from the neighbor (segregation), while solitary plants placed more roots towards the middle of the pot. Total biomass remained unaffected. Database comparisons in transcriptome analysis revealed considerable similarity between Arabidopsis root reactions to neighbors and reactions to pathogens. Detailed analyses of the functional category ‘biotic stress’ using MapMan tools found the sub-category ‘pathogenesis-related proteins’ highly significantly induced. A comparison to a study on intraspecific competition brought forward a core of genes consistently involved in reactions to neighbor roots. We conclude that beyond resource depletion roots perceive neighboring roots or their associated microorganisms by a relatively uniform mechanism that involves the strong induction of pathogenesis-related proteins. In an ecological context the findings reveal that belowground neighbor detection may occur independently of resource depletion, allowing for a time advantage for the root to prepare for potential interactions.

/pdf/belowground-neighbor-perception-in-arabidopsis-thaliana-ohzk7auz5s.pdf

Belowground neighbor perception in Arabidopsis thaliana studied by transcriptome analysis: roots of Hieracium pilosella cause biotic stress

In addition to the role of the cell wall as a physical barrier against pathogens, some of its constituents, such as pectin-derived oligogalacturonides (OGAs) are essential components to trigger signaling pathways that induce rapid defense responses. Many pathogens directly penetrate the cell wall to access water and nutrients of the plant protoplast, and a rigid cell wall can fend off pathogen attack by forming an impenetrable physical barrier. Thus, cell wall integrity sensing is one mechanism by which plants may detect pathogen attack. Moreover, when the plant-pathogen interaction occurred, OGAs released during cell wall modification can trigger plant defense (e.g., production of reactive oxygen species, production of anti-microbial metabolites and synthesis of pathogenesis-related proteins). This review documents and discusses studies suggesting that OGAs play a dual signaling role during pathogen attack by inducing defense responses and plant architecture adjustment.

https://www.tandfonline.com/doi/pdf/10.4161/psb.21779

Signaling role of oligogalacturonides derived during cell wall degradation

Phytoalexin accumulation is one of a myriad of plant defense responses; these responses can be elicited by pathogens or molecules such as oligogalacturonides (OGAs). Phytoalexin production has been considered a vital component of the resistance mechanisms that determine the outcome of many plant-microbe interactions. Besides inducing defense responses, OGAs have been shown to affect plant development, which normally is controlled by plant hormones, particularly auxin. In this work we measured phytoalexin accumulation in roots of bean (Phaseolus vulgaris L.) seedlings grown in the presence or absence of the auxin 3-naphtalenacetic acid (NAA) and treated with a decagalacturonide (OGA10). We found that OGA10 (0.01 mM) caused phytoalexin production and also inhibited main root elongation and the formation of secondary roots by ca. 33%. Expression of Cycb 2-2 was also inhibited, while pal and chs were highly expressed. The root growth inhibition was not overcome by the addition of a stimulatory concentration of auxin (NAA 0.1 µM). The data suggests that elicitation of defense responses in the root alters metabolism in such a way that results in the modification of the architecture of bean roots.

/pdf/the-architecture-of-phaseolus-vulgaris-root-is-altered-when-1yxh42n1zk.pdf

The architecture of Phaseolus vulgaris root is altered when a defense response is elicited by an oligogalacturonide

Measurement of cytosolic calcium concentration in protoplasts of bean leaf was carried out using Fluo-3/AM indicator. The uptake of the fluorescent indicator, as its permeable ester, was achieved by employing pluronic acid F-127 in the incubating solution. In the absence of pluronic acid F-127 loading was two times lower in the same time period. A rapid intracellular location of the fluorescent dye after intracellular hydrolysis is reported, with no vacuole trapping of the fluorescent indicator. An increase from 54 to 91 nM free calcium ([Ca2+]i) was observed immediately after addition of a pectic elicitor. Advantages of the use of pluronic acid F-127 are in the speed of the indicator entry, eliminating any possible hydrolysis by external esterases, and more efficient loading while maintaining cell integrity. © 1997 John Wiley & Sons, Ltd.

Use of pluronic acid F‐127 with Fluo‐3/AM probe to determine intracellular calcium changes elicited in bean protoplasts

Phytoalexins are secondary metabolites that in legumes are typically isoflavonoid derivatives. Techniques for their measurement very often use determination of the UV absorbance of ethanol extracts to avoid time-consuming extractions and the use of expensive solvents. However, other phenolic compounds also give UV absorbances interfering with quantitative determinations. A microassay is proposed to extract and measure bean phytoalexins from intact, viable, bean leaf protoplasts. Protoplasts as a tool for measuring phytoalexin elicitor stimulation is discussed.

Eva Soriano

Papers

The architecture of Phaseolus vulgaris root is altered when a defense response is elicited by an oligogalacturonide

Use of pluronic acid F‐127 with Fluo‐3/AM probe to determine intracellular calcium changes elicited in bean protoplasts

Microassay to measure phytoalexin response in protoplasts of bean leaves