Hydroponic cultivation conditions allowing the reproducible investigation of poplar root suberization and water transport

Background With increasing joint research cooperation on national and international levels, there is a high need for harmonized and reproducible cultivation conditions and experimental protocols in order to ensure the best comparability and reliability of acquired data. As a result, not only comparisons of findings of different laboratories working with the same species but also of entirely different species would be facilitated. As Populus is becoming an increasingly important genus in modern science and agroforestry, the integration of findings with previously gained knowledge of other crop species is of high significance. Results To ease and ensure the comparability of investigations of root suberization and water transport, on a high degree of methodological reproducibility, we set up a hydroponics-based experimental pipeline. This includes plant cultivation, root histochemistry, analytical investigation, and root water transport measurement. A 5-week-long hydroponic cultivation period including an optional final week of stress application resulted in a highly consistent poplar root development. The poplar roots were of conical geometry and exhibited a typical Casparian band development with subsequent continuously increasing suberization of the endodermis. Poplar root suberin was composed of the most frequently described suberin substance classes, but also high amounts of benzoic acid derivatives could be identified. Root transport physiology experiments revealed that poplar roots in this developmental stage have a two- to tenfold higher hydrostatic than osmotic hydraulic conductivity. Lastly, the hydroponic cultivation allowed the application of gradually defined osmotic stress conditions illustrating the precise adjustability of hydroponic experiments as well as the previously reported sensitivity of poplar plants to water deficits. Conclusions By maintaining a high degree of harmonization, we were able to compare our results to previously published data on root suberization and water transport of barley and other crop species. Regarding hydroponic poplar cultivation, we enabled high reliability, reproducibility, and comparability for future experiments. In contrast to abiotic stress conditions applied during axenic tissue culture cultivation, this experimental pipeline offers great advantages including the growth of roots in the dark, easy access to root systems before, during, and after stress conditions, and the more accurate definition of the developmental stages of the roots.

The mutualism effector MiSSP7 of Laccaria bicolor alters the interactions between the poplar JAZ6 protein and its associated proteins

Despite the pivotal role of jasmonic acid in the outcome of plant-microorganism interactions, JA-signaling components in roots of perennial trees like western balsam poplar (Populus trichocarpa) are poorly characterized. Here we decipher the poplar-root JA-perception complex centered on PtJAZ6, a co-repressor of JA-signaling targeted by the effector protein MiSSP7 from the ectomycorrhizal basidiomycete Laccaria bicolor during symbiotic development. Through protein–protein interaction studies in yeast we determined the poplar root proteins interacting with PtJAZ6. Moreover, we assessed via yeast triple-hybrid how the mutualistic effector MiSSP7 reshapes the association between PtJAZ6 and its partner proteins. In the absence of the symbiotic effector, PtJAZ6 interacts with the transcription factors PtMYC2s and PtJAM1.1. In addition, PtJAZ6 interacts with it-self and with other Populus JAZ proteins. Finally, MiSSP7 strengthens the binding of PtJAZ6 to PtMYC2.1 and antagonizes PtJAZ6 homo-/heterodimerization. We conclude that a symbiotic effector secreted by a mutualistic fungus may promote the symbiotic interaction through altered dynamics of a JA-signaling-associated protein–protein interaction network, maintaining the repression of PtMYC2.1-regulated genes.

Characteristics of Organic Acid Secretion Associated with the Interaction betweenBurkholderia multivoransWS-FJ9 and Poplar Root System

The objective of this study was to investigate whether plant-bacteria interaction affects the secretion of organic acids by both organisms and to assess whether the production of IAA by the bacterium increases the secretion of organic acids by root exudates, and if the stress produced by low available phosphorus (P) affects the production of organic acids by bacteria, by roots, or by root exudates in presence of bacterial cultures. With this purpose, we used as a biological model poplar plants and one strain ofBurkholderia multivoransable to solubilize P. High performance liquid chromatography was utilized to measure organic acids. The tests, the inductive effects of exogenous indole-3-acetic acid (IAA) on secretion of organic acids, the 2 × 4 × 2 factorial design experiment, and the ability of organic acids to solubilize tricalcium phosphate were performed to investigate the interactive effects. The results showed that, afterB. multivoransWS-FJ9 interacted with the poplar root system, the key phosphate-solubilizing driving force was gluconic acid (GA) which was produced in three ways: (1) secreted by the root system in the presence of IAA produced byB. multivoransWS-FJ9; (2) secreted byB. multivoransWS-FJ9; and (3) secreted by the poplar root system in the presence of phosphorus stress. When phosphorus stress was absent, the GA was produced as outlined in (1) and (2) above. These results demonstrated that inoculatingB. multivoransWS-FJ9 into the poplar root system could increase the amount of GA secretion and implied that the interaction betweenB. multivoransWS-FJ9 and the poplar root system could contribute to the increase of P available fraction for poplar plants.