Differential strategies of two species of arbuscular mycorrhizal fungi in the protection of maize plants grown in chromium-contaminated soils.

Chromium (Cr) is a nonessential element for plants that is extremely toxic at high concentrations. Zea mays L. is a species of plant that has developed adaptive mechanisms to increase its tolerance and absorption capacity for this metal. One effective mechanism is to form associations with arbuscular mycorrhizal fungi (AMF), which help the plant cope with stress from heavy metals such as Cr. However, it is still not clear which species of AMF are more efficient as bioremediating agents for plants of agricultural importance. Here, we evaluated the effect of Gigaspora gigantea and Rhizophagus irregularis as protective agents of maize plants in soils contaminated with Cr at concentrations of 0, 48.5, 97 and 194 mg kg-1 under greenhouse conditions. Maize tolerance to Cr was corroborated, as well as increased absorption of this element by plants associated with both species of AMF. G. gigantea caused an increase in nitrogen content and greater translocation of Cr to the aerial part of the plant; R. irregularis registered an increase in the formation of arbuscules and vesicles with increasing metal concentration and greater retention of Cr in the roots of the plants. Based on these results, we can conclude that the analyzed species of fungi use different strategies, with similar effectiveness, to enhance the absorption capacity of Cr by the plant and influence the differential deposition of this metal in various parts of the plant.

YUCCA4 overexpression modulates auxin biosynthesis and transport and influences plant growth and development via crosstalk with abscisic acid in Arabidopsis thaliana.

Auxin regulates a plethora of events during plant growth and development, acting in concert with other phytohormones. YUCCA genes encode flavin monooxygenases that function in tryptophan-dependent auxin biosynthesis. To understand the contribution of the YUCCA4 (YUC4) gene on auxin homeostasis, plant growth and interaction with abscisic acid (ABA) signaling, 35S::YUC4 seedlings were generated, which showed elongated hypocotyls with hyponastic leaves and changes in root system architecture that correlate with enhanced auxin responsive gene expression. Differential expression of PIN1, 2, 3 and 7 auxin transporters was detected in roots of YUC4 overexpressing seedlings compared to the wild-type: PIN1 was down-regulated whereas PIN2, PIN3 and PIN7 were up-regulated. Noteworthy, 35S::YUC4 lines showed enhanced sensitivity to ABA on seed germination and post-embryonic root growth, involving ABI4 transcription factor. The auxin reporter genes DR5::GUS, DR5::GFP and BA3::GUS further revealed that abscisic acid impairs auxin responses in 35S::YUC4 seedlings. Our results indicate that YUC4 overexpression influences several aspects of auxin homeostasis and reveal the critical roles of ABI4 during auxin-ABA interaction in germination and primary root growth.

Antibacterial and cytotoxicity activities and phytochemical analysis of three ornamental plants grown in Mexico.

This study evaluates the antibacterial, cytotoxic activities, and phytochemical composition, of Callistemon citrinus, Hibiscus rosa-sinensis and Plumbago auriculata leaves and flowers, three ornamental plants in Mexico. However, in other countries offers a range of other uses. Ethanol extracts of C. citrinus leaf and flower presented stronger antibacterial activity than the extracts obtained from the other two plants. C. citrinus leaf showed low cytotoxicity (LC50 <600 µg/mL) on the brine shrimp test, whereas the ethanol extracts of H. rosa-sinensis and P. auriculata leaves showed no cytotoxic activity. Flower extracts obtained from the three plants did no exhibit cytotoxicity. GC-MS analysis revealed that the ethanol extract of P. auriculata leaf contained lupeol triterpene and lupeol acetate, neither of them have been previously reported in this genus. Gamma sitosterol was present in the leaf and flower extracts of P. auriculata. Higher contents of linoleic and linolenic acids were found in extracts of H. rosa-sinensis leaves and flowers. The ability of the ethanol extracts of C. citrinus leaves and flowers to inhibit the growth of Gram-positive and Gram-negative bacteria indicates a potentially broad antimicrobial spectrum. Moreover, the absence of cytotoxicity suggests the potential use of this plant to treat microbial infections.

Expression analysis of the Arabidopsis thaliana AtSpen2 gene, and its relationship with other plant genes encoding Spen proteins

Abstract Proteins of the Split ends (Spen) family are characterized by an N-terminal domain, with one or more RNA recognition motifs and a SPOC domain. In Arabidopsis thaliana, the Spen protein FPA is involved in the control of flowering time as a component of an autonomous pathway independent of photoperiod. The A. thaliana genome encodes another gene for a putative Spen protein at the locus At4g12640, herein named AtSpen2. Bioinformatics analysis of the AtSPEN2 SPOC domain revealed low sequence similarity with the FPA SPOC domain, which was markedly lower than that found in other Spen proteins from unrelated plant species. To provide experimental information about the function of AtSpen2, A. thaliana plants were transformed with gene constructs of its promoter region with uidA::gfp reporter genes; the expression was observed in vascular tissues of leaves and roots, as well as in ovules and developing embryos. There was absence of a notable phenotype in knockout and overexpressing lines, suggesting that its function in plants might be specific to certain endogenous or environmental conditions. Our results suggest that the function of Atspen2 diverged from that of fpa due in part to their different transcription expression pattern and divergence of the regulatory SPOC domain.

Expression analysis of the Arabidopsis thaliana AtSpen2 gene, and its relationship with other plant genes encoding Spen proteins.

Proteins of the Split ends (Spen) family are characterized by an N-terminal domain, with one or more RNA recognition motifs and a SPOC domain. In Arabidopsis thaliana, the Spen protein FPA is involved in the control of flowering time as a component of an autonomous pathway independent of photoperiod. The A. thaliana genome encodes another gene for a putative Spen protein at the locus At4g12640, herein named AtSpen2. Bioinformatics analysis of the AtSPEN2 SPOC domain revealed low sequence similarity with the FPA SPOC domain, which was markedly lower than that found in other Spen proteins from unrelated plant species. To provide experimental information about the function of AtSpen2, A. thaliana plants were transformed with gene constructs of its promoter region with uidA::gfp reporter genes; the expression was observed in vascular tissues of leaves and roots, as well as in ovules and developing embryos. There was absence of a notable phenotype in knockout and overexpressing lines, suggesting that its function in plants might be specific to certain endogenous or environmental conditions. Our results suggest that the function of Atspen2 diverged from that of fpa due in part to their different transcription expression pattern and divergence of the regulatory SPOC domain.

Arabidopsis thaliana sucrose phosphate synthase (sps) genes are expressed differentially in organs and tissues, and their transcription is regulated by osmotic stress.

Sucrose is synthesized from UDP-Glc and Fru-6-phosphate via the activity of sucrose-phosphate synthase (SPS) enzymes, which produce Suc-6-phosphate. Suc-6-phosphate is rapidly dephosphorylated by phosphatases to produce Suc and inorganic phosphate. Arabidopsis has four sps genes encoding SPS enzymes. Of these enzymes, AtSPS1F and AtSPS2F have been grouped with other dicotyledonous SPS enzymes, while AtSPS3F and AtSPS4F are included in groups with both dicotyledonous and monocotyledonous SPS enzymes. In this work, we generated Arabidopsis thaliana transformants containing the promoter region of each sps gene fused to gfp::uidA reporter genes. A detailed characterization of expression conferred by the sps promoters in organs and tissues was performed. We observed expression of AtSPS1F, AtSPS2F and AtSPS3F in the columella roots of the plants that support sucrose synthesis. Hence, these findings support the idea that sucrose synthesis occurs in the columella cells, and suggests that sucrose has a role in this tissue. In addition, the expression of AtSPS4F was identified in embryos and suggests its participation in this developmental stage. Quantitative transcriptional analysis of A. thaliana plants grown in media with different osmotic potential showed that AtSPS2F and AtSPS4F respond to osmotic stress.

Arbuscular mycorrhizal symbiosis increases the content of volatile terpenes and plant performance in Satureja macrostema (Benth. ) Briq / La simbiosis micorrícica arbuscular aumenta el contenido de terpenos volátiles y el rendimiento vegetal en Satureja macrostema (Benth.) Briq

We studied the effect of Rhizophagus irregularis on plant performance and volatile terpenes content of the Mexican native medicinal plant Satureja macrostema (Benth.) Briq. (Lamiaceae) in greenhouse conditions. The growth parameters considered in this research and the composition of volatile components were quantified monthly in mycorrhizal and non-mycorrhizal plants. The essential oil was collected from aerial parts and analyzed by gas chromatography-mass spectrometry. Colonization by R. irregularis significantly increased biomass, shoot and root length, and the amount of volatile terpenes. The more concentrated volatile terpenes were limonene, β- linalool, menthone, pulegone, and verbenol acetate. It is concluded that the use of R. irregularis allows optimal growth of S. macrostema plants in low fertility soils and increased production of the main components of the essential oil.

El efecto de Rhizophagus irregularis sobre el rendimiento vegetal y la producción de los terpenos volátiles de Satureja macrostema (Benth.) Briq. (Lamiaceae), una planta medicinal nativa mexicana, fue estudiado en condiciones de invernadero. Los parámetros de crecimiento considerados en esta investigación y los componentes volátiles, fueron cuantificados mensualmente en plantas con y sin micorrizas. El aceite esencial fue colectado de la parte aérea y fue analizado por técnicas de cromatografía de gases-espectrometría de masas. La colonización de R. irregularis aumentó significativamente la biomasa, longitud de tallo y raíz, y la cantidad de terpenos volátiles. Los terpenos volátiles mayoritarios fueron limoneno, β-linalol, mentona, pulegona y acetato de verbenol. Se concluye que el uso de R. irregularis permitió un óptimo crecimiento de las plantas de S. macrostema en suelos de baja fertilidad, con un aumento de los componentes principales del aceite esencial.

Effect of mineral nutrients on the uptake of Cr(VI) by maize plants.

To determine the potential of maize plants for phytoextraction of chromium (Cr), and the effect that some mineral nutrients have on this process, the uptake of this metal was analyzed in vitro. 12-day-old plants were incubated in nutrient solutions with 200 or 250 µM potassium chromate and 3mM supplements of nitrate, phosphate or sulfate. The greatest accumulation of Cr after 48 hours was in plants that were supplemented with nitrate (800 and 350 mg kg(-1) DW for roots and stems, respectively) with bioaccumulation factors of 18 and 7 in the roots and in the foliage, respectively. Moreover, the translocation factor from root to leaves was 0.4 when nitrate was supplemented; thus, the addition of this nutrient is recommended when the crop is to be used for phytoextraction of Cr. Considering the results obtained, the high biomass produced by this crop and the extensive knowledge of agricultural practices for this plant, we believe that maize has the potential to be considered in the phytoextraction of Cr, based on preliminary assays of soil conditions.

Chromate induces adventitious root formation via auxin signalling and SOLITARY-ROOT/IAA14 gene function in Arabidopsis thaliana.

Morphological root plasticity optimizes nutrient and water uptake by plants and is a promising target to improve tolerance to metal toxicity. Exposure to sublethal chromate [Cr(VI)] concentrations inhibits root growth, decreases photosynthesis and compromises plant development and productivity. Despite the increasing environmental problem that Cr(VI) represents, to date, the Cr tolerance mechanisms of plants are not well understood, and it remains to be investigated whether root architecture remodelling is important for plant adaptation to Cr(VI) stress. In this report, we analysed the growth response of Arabidopsis thaliana seedlings to concentrations of Cr(VI) that strongly repress primary and lateral root growth. Interestingly, adventitious roots started developing, branched and allowed seedlings to grow under highly growth-repressing Cr(VI) concentrations. Cr(VI) negatively regulates auxin transport and response gene expression in the primary root tip, as evidenced by decreased expression of auxin-related reporters DR5::GFP, DR5::uidA and PIN1::PIN1::GFP, and then, another auxin maximum is established at the site of adventitious root initiation that drives adventitious root organogenesis. Both primary root growth inhibition and adventitious root formation induced by high Cr(VI) levels are blocked by a gain-of-function mutation in the SOLITARY-ROOT/IAA14 gene of Arabidopsis. These data provide evidence that suggests a critical role for auxin transport and signalling via IAA14/SLR1 in the developmental program linking Cr(VI) to root architecture remodelling.

Phosphate relieves chromium toxicity in Arabidopsis thaliana plants by interfering with chromate uptake.

Soil contamination by hexavalent chromium [Cr(VI) or chromate] due to anthropogenic activities has become an increasingly important environmental problem. Mineral nutrients such as phosphate (Pi), sulfate and nitrate have been reported to attenuate Cr(VI) toxicity, but the underlying mechanisms remain to be clarified. Here, we show that chromate activates the expression of low-Pi inducible reporter genes AtPT1 and AtPT2 in Arabidopsis thaliana transgenic seedlings. Primary-root growth was inhibited by 60 % in AtPT2::uidA-expressing seedlings upon exposure to 140-µM Cr(VI). However, increasing the Pi and sulfate supply to the seedlings that were experiencing Cr(VI) toxicity completely and partially restored the root growth, respectively. This effect correlated with the Cr(VI)-induced AtPT2::uidA expression being completely reversed by addition of Pi. To evaluate whether the nutrient supply may affect the endogenous level of Cr in plants grown under toxic Cr(VI) levels, the contents of Cr were measured (by ICP-MS analyses) in seedlings treated with Cr and with or without Pi, sulfate or nitrate. It was found that Cr accumulation increases tenfold in plants treated with 140-µM Cr(VI) without modifying the phosphorus concentration in the plant. In contrast, the supply of Pi specifically decreased the Cr content to levels similar to those found in seedlings grown in medium without chromate. Taken together, these results show that in A. thaliana seedlings the uptake of Cr(VI) is reduced by Pi. Moreover, our data indicate that Pi and sulfate supplements may be useful in strategies for handling Cr-contaminated soils.