Gabriela Sepúlveda-Jiménez

Nitrogen deficiency stimulates biosynthesis of bioactive phenylethanoid glycosides in the medicinal plant Castilleja tenuiflora Benth.

AbstractCastilleja tenuiflora Benth. (Orobanchaceae) in vitro cultures are alternative sources of phenylethanoid glycosides (PhGs), promising natural products for chronic diseases treatment because of their extensive range of biological activities. To increase the yields of these bioactive compounds, the identification of factors affecting their biosynthesis is required. Here, we show that N deficiency stimulates PhGs biosynthesis by increasing the activity of PAL. The contents of two PhGs, verbascoside and isoverbascoside, were enhanced under N deficiency compared with control conditions. The maximum PAL activity in shoots cultured under N deficiency was also increased. Furthermore, we found de novo anthocyanin synthesis under N deficiency suggesting an additional impact of this stress factor on flavonoid metabolism. N deficiency negatively affected the shoot growth (length, biomass, and total chlorophyll content) and multiplication rate, and inhibited the root formation of C. tenuiflora shoots. Our results demonstrate that N deficiency increases PhGs biosynthesis in C. tenuiflora.

Induction of resistance to Sclerotium rolfsii in different varieties of onion by inoculation with Trichoderma asperellum

We evaluated the ability of Trichoderma asperellum Samuels, Lieckfeldt & Nirenberg to induce resistance to the fungal plant pathogen, Sclerotium rolfsii Saccardo, in three onion (Allium cepa L.) varieties. Both the severity of disease and the activities of glucanase, chitinase and peroxidase (enzymes involved in plant resistance) were evaluated in onions inoculated with T. asperellum alone, S. rolfsii alone, or both T. asperellum and S. rolfsii (dual-inoculation) and compared to uninoculated (control) plants. In dual inoculations, the presence of T. asperellum reduced the severity of disease symptoms caused by S. rolfsii. Inoculation with T. asperellum alone increased glucanase, chitinase and peroxidase activity in bulbs, roots and leaves of all three onion varieties compared to uninoculated controls; bulbs of the variety Red Satan (RS) had the highest enzyme activity. In plants inoculated with S. rolfsii alone, enzyme activity was only increased in bulbs and roots compared to uninoculated controls. The highest levels of enzyme activity also occurred only in bulbs and roots of plants that had been dual-inoculated with T. asperellum and S. rolfsii. Plants of the RS variety showed the highest enzyme activities (both constitutive and induced) and showed the lowest severity of disease. Therefore, application of T. asperellum has potential as a biological control alternative to synthetic fungicides for protection of onion crops against infection by S. rolfsii. This protection depends on both constitutive and induced defence responses and varies amongst onion varieties.

Robust root growth in altered hydrotropic response1 (ahr1) mutant of Arabidopsis is maintained by high rate of cell production at low water potential gradient.

Hydrotropism is the directional root growth response determined by water stimulus. In a water potential gradient system (WPGS) the roots of the Arabidopsis wild type have a diminished root growth compared to normal medium (NM). In contrast, the altered hydrotropic response1 (ahr1) mutant roots maintain their robust growth in the same WPGS. The aims of this work were to ascertain how ahr1 roots could sustain growth in the WPGS, with a special focus on the integration of cellular processes involved in the signaling that determines root growth during abiotic stress and their relation to hydrotropism. Cellular analysis of the root apical meristem of ahr1 mutant contrary to the wild type showed an absence of changes in the meristem length, the elongation zone length, the length of fully elongated cells, and the cell cycle duration. The robust and steady root growth of ahr1 seedlings in the WPGS is explained by the mutant capacity to maintain cell production and cell elongation at the same level as in the NM. Analysis of auxin response at a transcriptional level showed that roots of the ahr1 mutant had a lower auxin response when grown in the WPGS, compared to wild type, indicating that auxin signaling participates in attenuation of root growth under water stress conditions. Also, wild type plants exhibited a high increase in proline content while ahr1 mutants showed minimum changes in the Normal Medium→Water Stress Medium (NM→WSM), a lower water potential gradient system than the WPGS. Accordingly, in this condition, gene expression of Δ1-6 Pyrroline-5-Carboxylate Synthetase1 (P5CS1) involved in proline synthesis strongly increased in wild type but not in ahr1 seedlings. The ahr1 phenotype shows unique features since the mutant root cells continue to proliferate and grow in the presence of a progressively negative water potential gradient at a level comparable to wild type growing in the NM. As such, it represents an exceptional resource for understanding hydrotropism.

Autophagy mediates hydrotropic response in Arabidopsis thaliana roots

This work shows that autophagy plays a key role in the hydrotropic curvature of Arabidopsis thaliana roots. An analysis of GFP-ATG8a transgenic plants showed that autophagosomes accumulated in the root curvature 2 h after the transfer of seedlings to Normal Medium-Water Stress Medium (NM-WSM). Autophagy flux was required for root bending. Remarkably, several atg mutants did not show hydrotropic curvature in NM-WSM or the splitting-agar system. Hyper, an H2O2 sensor showed that H2O2 preferentially accumulated in the root curvature at a similar rate as the autophagosomes did during hydrotropic response. Peroxidase and ROBH activity inhibition affected, negatively or positively root curvature. This data suggested H2O2 balance was required for root bending. Malondialdehyde, a metabolite used as an indicator of oxidative stress, accumulated at the same rate during the development of the curvature in NM-WSM. These results suggest that autophagy is required for the hydrotropic response in NM-WSM. We discuss the possible regulatory role of H2O2 on autophagy during the hydrotropic response that might relieve oxidative stress provoked by water stress. NM-WSM is water stress system suitable for studying hydrotropic responses on a short-term basis.