We report on the homo-and hetero-transglycosylation activities of the HvXET3 and HvXET4 xyloglucan xyloglucosyl transferases (XET; EC 2.4.1.207) from barley (Hordeum vulgare L.), and the visualisation of these activities in young barley roots using Alexa Fluor 488-labelled oligosaccharides. We discover that these isozymes catalyse the transglycosylation reactions with the chemically defined donor and acceptor substrates, specifically with the xyloglucan donor and the penta-galacturonide [a(1-4)GalAp] 5 acceptorthe homogalacturonan (pectin) fragment. This activity is supported by 3D molecular models of HvXET3 and HvXET4 with the docked XXXG donor and [a(1-4)GalAp] 5 acceptor substrates at the À4 to +5 subsites in the active sites. Comparative sequence analyses of barley isoforms and seed-localised TmXET6.3 from nasturtium (Tropaeolum majus L.) permitted the engineering of mutants of TmXET6.3 that could catalyse the hetero-transglycosylation reaction with the xyloglucan/[a(1-4)GalAp] 5 substrate pair, while wild-type TmXET6.3 lacked this activity. Expression data obtained by real-time quantitative polymerase chain reaction of HvXET transcripts and a clustered heatmap of expression profiles of the gene family revealed that HvXET3 and HvXET6 co-expressed but did not share the monophyletic origin. Conversely, HvXET3 and HvXET4 shared this relationship, when we examined the evolutionary history of 419 glycoside hydrolase 16 family members, spanning monocots, eudicots and a basal Angiosperm. The discovered hetero-transglycosylation activity in HvXET3 and HvXET4 with the xyloglucan/[a(1-4)GalAp] 5 substrate pair is discussed against the background of roles of xyloglucan-pectin heteropolymers and how they may participate in spatial patterns of cell wall formation and remodelling , and affect the structural features of walls.

No abstract

Auxins are plant hormones that affect plant growth, development, and improve a plant’s tolerance to stress. In this study, we found that the application of indole-3-butyric acid (IBA) had diverse effects on the growth of maize (Zea mays L.) roots treated without/with Cd. IBA caused changes in the growth and morphology of the roots under non-stress conditions; hence, we were able to select two concentrations of IBA (10−11 M as stimulatory and 10−7 M as inhibitory). IBA in stimulatory concentration did not affect the concentration of H2O2 or the activity of antioxidant enzymes while IBA in inhibitory concentration increased only the concentration of H2O2 (40.6%). The application of IBA also affected the concentrations of mineral nutrients. IBA in stimulatory concentration increased the concentration of N, K, Ca, S, and Zn (5.8–14.8%) and in inhibitory concentration decreased concentration of P, K, Ca, S, Fe, Mn, Zn, and Cu (5.5–36.6%). Moreover, IBA in the concentration 10−9 M had the most positive effects on the plants cultivated with Cd. It decreased the concentration of H2O2 (34.3%), the activity of antioxidant enzymes (23.7–36.4%), and increased the concentration of all followed elements, except Mg (5.5–34.1%), when compared to the Cd.