Microbacterium plantarum sp. nov. and Microbacterium thalli sp. nov., two endophytic metal-resistant bacteria isolated from Sphaeralcea angustifolia (Cav.) G. Don and Prosopis laevigata (Humb. et Bonpl. ex Willd) M.C. Johnston.

Four Gram-positive, aerobic, catalase- and oxidase-negative, rod-shaped, motile endophytic bacterial strains, designated NM3R9T, NE1TT3, NE2TL11 and NE2HP2T, were isolated from the inner tissues (leaf and stem) of Sphaeralcea angustifolia and roots of Prosopis laevigata. They were characterized using a polyphasic approach, which revealed that they represent two novel Microbacterium species. Phylogenetic analysis based on 16S rRNA gene sequencing showed that the species closest to NE2HP2T was Microbacterium arborescens DSM 20754T (99.6 %) and that closest to NM3R9T, NE2TL11 and NE2TT3 was Microbacterium oleivorans NBRC 103075T (97.4 %). The whole-genome average nucleotide identity value between strain NM3R9T and Microbacterium imperiale DSM 20530T was 90.91 %, and that between strain NE2HP2T and M. arborecens DSM 20754T was 91.03 %. Digital DNA-DNA hybridization showed values of less than 70 % with the type strains of related species. The polar lipids present in both strains included diphosphatidylglycerol, phosphatidylglycerol, glycolipids and unidentified lipids, whereas the major fatty acids included anteiso-C15 : 0, anteiso-C17 : 0, iso-C16 : 0 and C16 : 0. Whole-cell sugars included mannose, rhamnose and galactose. Strains NM3R9T and NE2HP2T showed physiological characteristics different from those present in closely related Microbacterium species. According to the taxonomic analysis, both strains belong to two novel species. The name Microbacterium plantarum sp. nov. is proposed for strain NE2HP2T (=LMG 30875T=CCBAU 101117T) and Microbacterium thalli sp. nov. for strains NM3R9T (=LMG 30873T=CCBAU 101116T), NE1TT3 (=CCBAU 101114) and NE2TL11 (=CCBAU 101115).

Phytochemical Analysis and Biological Activities of Ripe Fruits of Mistletoe (Psittacanthus calyculatus).

Psittacanthus calyculatus is a hemiparasitic plant of an arboreal species (e.g., forest, fruit trees). Its foliage has therapeutic potential; however, little is known about its fruits. In this study, the phytochemical profile and biological activities of P. calyculatus fruits hosted by Prosopis laevigata and Quercus deserticola were evaluated. The fruits of P. calyculatus from P. laevigata showed the highest content of total phenols (71.396 ± 0.676 mg GAE/g DW). The highest content of flavonoids and anthocyanins was presented in those from Q. deserticola (14.232 ± 0.772 mg QE/g DW; 2.431 ± 0.020 mg C3GE/g DW). The anthocyanin cyanidin-3-glucoside was detected and quantified via high-performance thin-layer chromatography (HPTLC) (306.682 ± 11.804 mg C3GE/g DW). Acidified extracts from host P. laevigata showed the highest antioxidant activity via ABTS•+ (2,2'azinobis-(3-ethylbenzothiazdin-6-sulfonic acid) (214.810 ± 0.0802 mg TE/g DW). Fruit extracts with absolute ethanol from the P. laevigata host showed the highest antihypertensive activity (92 ± 3.054% inhibition of an angiotensin converting enzyme (ACE)). Fruit extracts from both hosts showed a minimum inhibitory concentration (MIC) of 6.25 mg/mL and a minimum bactericidal concentration (MBC) of 12.5 mg/mL against Escherichia coli, Salmonella choleraesuis and Shigella flexneri. Interestingly, a significant host effect was found. P. calyculatus fruits extract could be used therapeutically. However, further confirmation experiments should be carried out.

Aspergillus luchuensis, an Endophyte Fungus from the Metal Hyperaccumulator Plant Prosopis laevigata, Promotes Its Growth and Increases Metal Translocation.

Heavy metal pollution is a worldwide environmental and human health problem. Prosopis laevigata is a hyperaccumulator legume that bioaccumulates Pb, Cu and Zn. With interest in designing phytoremediation strategies for sites contaminated with heavy metals, we isolated and characterized endophytic fungi from the roots of P. laevigata growing on mine tailings located in Morelos, Mexico. Ten endophytic isolates were selected by morphological discrimination and a preliminary minimum inhibitory concentration was determined for zinc, lead and copper. A novel strain of Aspergillus closest to Aspergillus luchuensis was determined to be a metallophile and presented a marked tolerance to high concentrations of Cu, Zn and Pb, so it was further investigated for removal of metals and promotion of plant growth under greenhouse conditions. The control substrate with fungi promoted larger size characters in P. laevigata individuals in comparison with the other treatments, demonstrating that A. luchuensis strain C7 is a growth-promoting agent for P. laevigata individuals. The fungus favors the translocation of metals from roots to leaves in P. laevigata, promoting an increased Cu translocation. This new A. luchuensis strain showed endophytic character and plant growth-promotion activity, high metal tolerance, and an ability to increase copper translocation. We propose it as a novel, effective and sustainable bioremediation strategy for copper-polluted soils.

Protein Profiling of Psittacanthus calyculatus during Mesquite Infection.

Psittacanthus calyculatus is a hemiparasite mistletoe that represents an ecological problem due to the impacts caused to various tree species of ecological and commercial interest. Although the life cycle for the Psittacanthus genus is well established in the literature, the development stages and molecular mechanism implicated in P. calyculatus host infection are poorly understood. In this study, we used a manageable infestation of P. laevigata with P. calyculatus to clearly trace the infection, which allowed us to describe five phenological infective stages of mistletoe on host tree branches: mature seed (T1), holdfast formation (T2), haustorium activation (T3), haustorium penetration (T4), and haustorium connection (T5) with the host tree. Proteomic analyses revealed proteins with a different accumulation and cellular processes in infective stages. Activities of the cell wall-degrading enzymes cellulase and ß-1,4-glucosidase were primarily active in haustorium development (T3), while xylanase, endo-glucanase, and peptidase were highly active in the haustorium penetration (T4) and xylem connection (T5). Patterns of auxins and cytokinin showed spatial concentrations in infective stages and moreover were involved in haustorium development. These results are the first evidence of proteins, cell wall-degrading enzymes, and phytohormones that are involved in early infection for the Psittacanthus genus, and thus represent a general infection mechanism for other mistletoe species. These results could help to understand the molecular dialogue in the establishment of P. calyculatus parasitism.

Assisted Phytostabilization of Mine-Tailings with Prosopis laevigata (Fabaceae) and Biochar.

Phytoremediation is a cost-effective technique to remediate heavy metal (HM) polluted sites. However, the toxic effects of HM can limit plant establishment and development, reducing phytoremediation effectiveness. Therefore, the addition of organic amendments to mine wastes, such as biochar, improves the establishment of plants and reduces the bioavailability of toxic HM and its subsequent absorption by plants. Prosopis laevigata can establish naturally in mine tailings and accumulate different HM; however, these individuals show morphological and genetic damage. In this study, the effect of biochar on HM bioaccumulation in roots and aerial tissues, HM translocation, morphological characters and plant growth were evaluated, after three and six months of exposure. Plants grown on mine tailings with biochar presented significantly higher values for most of the evaluated characters, in respect to plants that grew on mine tailing substrate. Biochar addition reduced the bioaccumulation and translocation of Cu, Pb, and Cd, while it favored the translocation of essential metals such as Fe and Mn. The addition of biochar from agro-industrial residues to mine tailings improves the establishment of plants with potential to phytoextract and phytostabilize metals from polluted soils. Using biochar and heavy metal accumulating plants constitutes an assisted phytostabilization strategy with great potential for HM polluted sites such as Cd and Pb.

Long-Chain Hydrocarbons (C21, C24, and C31) Released by Bacillus sp. MH778713 Break Dormancy of Mesquite Seeds Subjected to Chromium Stress.

Volatile organic compounds (VOCs) produced by rhizobacteria have been proven to stimulate plant growth during germination and seedling stages. However, the modulating effect of bacterial volatiles on the germination of seeds subjected to heavy metal stress is scarcely studied. In this work, the ability of volatiles released by Bacillus sp. MH778713 to induce seed dormancy breakage in Prosopis laevigata and Arabidopsis thaliana seeds were examined. The minimal inhibitory concentration of chromium (Cr) VI that prevents seed germination of P. laevigata and A. thaliana on water-Cr-agar plates was 2500 and 100 mg L-1, respectively. Remarkably, partitioned Petri-dish co-cultivation of Bacillus sp. MH778713 and plant seeds under Cr-stress showed the beneficial effect of volatiles emitted by Bacillus sp. MH778713, helping plant seeds to overcome Cr-stress. Among the metabolites emitted by Bacillus sp. MH778713, octadecane, heneicosane, 2,4-di-tert-butylphenol, hexadecane, eicosane, octacosane, and tetratriacontane were the most abundant. To confirm that these long-chain compounds produced by Bacillus sp. MH778713 could be responsible for the seed dormancy breakage, high pure organic compounds (2,4-di-tert-butylphenol, heneicosane, hentriacontane, and tetracosane) were used directly in germination assays of P. laevigata and A. thaliana seeds instead of volatiles emitted by Bacillus sp. MH778713. All organic compounds allowed Prosopis and Arabidopsis seeds to overcome Cr-toxicity and germinate. The results of this study provide new insight into the role of long-chain bacterial compounds produced by Bacillus sp. MH778713 as triggers of seed abiotic stress tolerance, surmounting chromium stress and stimulating seedling development.

Nutritional Characterization of Prosopis laevigata Legume Tree (Mesquite) Seed Flour and the Effect of Extrusion Cooking on its Bioactive Components.

Mesquite (Prosopis laevigata) is a legume tree widely distributed in Aridoamerica. The mature fruit of this legume is a pod, which is currently underutilized and has high nutritional potential. In the present work, mesquite seed flour is described in terms of its nutritional value, as well as the effect of extrusion cooking on its bioactive components. Mesquite seed flour is rich in fiber (7.73 g/100 g) and protein (36.51 g/100 g), with valine as the only limiting amino acid. Total phenolic compound contents in raw and extruded seed flour were 6.68 and 6.46 mg of gallic acid equivalents/g (mg GAE/g), respectively. 2-2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity values in raw and extruded seed flour were 9.11 and 9.32 mg of ascorbic acid equivalent/g (mg AAE/g), respectively. The absorbance at 290 nm, as an indicator of generation of Maillard reaction product (MRP), was the same for raw and extruded samples. Apigenin was the only flavonoid found in mesquite seed flour (41.6 mg/kg) and was stable in the extrusion process. The water absorption index (WAI) and water solubility index (WSI) were changed significantly during extrusion. The expansion of mesquite seed flour extrudates was null due to the high protein and fiber content in the sample. Extrusion cooking of mesquite seed flour is a useful form of technology for the industrialization of this underutilized and nutritionally valuable legume.

Nuevos recursos naturales como complemento de la dieta de caprinos durante la época seca, en el Valle de Tehuacán, México / New natural resources as diet supplement for domestic goats during the dry season, in the Tehuacán Valley, Mexico / Novos recursos naturais como complemento da dieta de caprinos durante a época seca, no Vale de Tehuacán, México

La ganadería caprina es una de las actividades más importantes para la subsistencia campesina en el Valle de Tehuacán, México Aquí, como en otras tantas zonas semiáridas del trópico, esta práctica enfrenta una marcada limitación debido a la escasez de forraje durante la época de sequía. Para evaluar las características nutricionales de la dieta del ganado caprino durante dicha época, se determinaron los principales componentes de la misma y sus características. Por otro lado, se evaluaron diferentes recursos vegetales de la zona, determinando sus propiedades bromatológicas con el objetivo de analizar su uso potencial como complemento alimenticio. Durante la época de sequía se observó una marcada limitación de forraje que obliga al ganado a consumir plantas con bajo contenido nutricional y alto contenido en metabolitos secundarios, produciendo una pérdida importante de peso. En conjunto, la dieta durante la época de sequía es deficitaria en proteína y fósforo. Los resultados indican que especies de Opuntia, Agave salmiana, los frutos de Yucca periculosa y las vainas de Prosopis laevigata y Acacia subangulata tienen un alto potencial para ser usadas, combinadas con el tradicional rastrojo de maíz, como complemento alimenticio. Estas nuevas fuentes de alimento posibilitan la estabulación o semi-estabulación del ganado durante la época de secas, como una alternativa de manejo. Ello reduciría la pérdida de energía en busca de recursos alimenticios, mejorando la productividad y disminuyendo el impacto sobre una de las vegetaciones más biodiversas de Norteamérica.

The extensive herding of goats has become the most important productive activity of subsistence for farmers at the Tehuacan Valley, Mexico. However, as in other semiarid tropical areas, the productivity of goat ranches is very low because of a very adverse period during the dry season. In order to determine the nutritional characteristics of the goats’ diet during this season, the diet was examined and the chemical characteristics of the principal species that are consumed was analyzed. On the other hand, different plant resources in the area, which can potentially be used as food supplementation, were analyzed determining their nutritional properties. A marked limitation of fodder during the dry season was found, which forces goats to eat plants with low nutritional characteristics and high content of secondary metabolites, provoking significant weight loss. Overall, grazing diet is deficient in protein and phosphorus during the dry season. The results indicate that species of Opuntia, Agave salmiana, the fruits of Yucca periculosa, and pods of Prosopis laevigata and Acacia subangulata present a high potential for use as food supplements, combined with the traditional corn stubble,. These new sources of nutrients make possible the enclosure of goats during the dry period, avoiding the high use of energy for food search, improving goat production and reducing goat impact on one of the most biodiverse area of vegetation North America.

A criação de gado caprino é uma das atividades mais importantes para a subsistência camponesa no Vale de Tehuacán, México. Aqui, como em outras tantas zonas semi-áridas do trópico, esta prática enfrenta uma marcada limitação devido à escassez de forragem durante a época de seca. Para avaliar as características nutricionais da dieta do gado caprino durante dita época, se determinaram os principais componentes da mesma e suas características. Por outro lado, se avaliaram diferentes recursos vegetais da zona, determinando suas propriedades bromatológicas com o objetivo de analisar seu uso potencial como complemento alimentício. Durante a época de seca se observou uma marcada limitação de forragem que obriga ao gado a consumir plantas com baixo conteúdo nutricional e alto conteúdo em metabólitos secundários, produzindo uma perda importante de peso. Em conjunto, a dieta durante a época de seca é deficitária em proteína e fósforo. Os resultados indicam que espécies de Opuntia, Agave salmiana, os frutos de Yucca periculosa e as vagens de Prosopis laevigata e Acacia subangulata têm um alto potencial para o uso combinado com o tradicional resíduo cultural do milho, como complemento alimentício. Estas novas fontes de alimento possibilitam a estabulação ou semi-estabulação do gado durante a época de secas, como uma alternativa de manejo. Isto reduziria a perda de energia em busca de recursos alimentícios, melhorando a produtividade e diminuindo o impacto sobre uma das vegetações mais biodiversas de Norte América.