The avocado genome informs deep angiosperm phylogeny, highlights introgressive hybridization, and reveals pathogen-influenced gene space adaptation.

The avocado, Persea americana, is a fruit crop of immense importance to Mexican agriculture with an increasing demand worldwide. Avocado lies in the anciently diverged magnoliid clade of angiosperms, which has a controversial phylogenetic position relative to eudicots and monocots. We sequenced the nuclear genomes of the Mexican avocado race, P. americana var. drymifolia, and the most commercially popular hybrid cultivar, Hass, and anchored the latter to chromosomes using a genetic map. Resequencing of Guatemalan and West Indian varieties revealed that ∼39% of the Hass genome represents Guatemalan source regions introgressed into a Mexican race background. Some introgressed blocks are extremely large, consistent with the recent origin of the cultivar. The avocado lineage experienced 2 lineage-specific polyploidy events during its evolutionary history. Although gene-tree/species-tree phylogenomic results are inconclusive, syntenic ortholog distances to other species place avocado as sister to the enormous monocot and eudicot lineages combined. Duplicate genes descending from polyploidy augmented the transcription factor diversity of avocado, while tandem duplicates enhanced the secondary metabolism of the species. Phenylpropanoid biosynthesis, known to be elicited by Colletotrichum (anthracnose) pathogen infection in avocado, is one enriched function among tandems. Furthermore, transcriptome data show that tandem duplicates are significantly up- and down-regulated in response to anthracnose infection, whereas polyploid duplicates are not, supporting the general view that collections of tandem duplicates contribute evolutionarily recent "tuning knobs" in the genome adaptive landscapes of given species.

An improved, low-cost, hydroponic system for growing Arabidopsis and other plant species under aseptic conditions.

BACKGROUND: Hydroponics is a plant growth system that provides a more precise control of growth media composition. Several hydroponic systems have been reported for Arabidopsis and other model plants. The ease of system set up, cost of the growth system and flexibility to characterize and harvest plant material are features continually improved in new hydroponic system reported. RESULTS: We developed a hydroponic culture system for Arabidopsis and other model plants. This low cost, proficient, and novel system is based on recyclable and sterilizable plastic containers, which are readily available from local suppliers. Our system allows a large-scale manipulation of seedlings. It adapts to different growing treatments and has an extended growth window until adult plants are established. The novel seed-holder also facilitates the transfer and harvest of seedlings. Here we report the use of our hydroponic system to analyze transcriptomic responses of Arabidopsis to nutriment availability and plant/pathogen interactions. CONCLUSIONS: The efficiency and functionality of our proposed hydroponic system is demonstrated in nutrient deficiency and pathogenesis experiments. Hydroponically grown Arabidopsis seedlings under long-time inorganic phosphate (Pi) deficiency showed typical changes in root architecture and high expression of marker genes involved in signaling and Pi recycling. Genome-wide transcriptional analysis of gene expression of Arabidopsis roots depleted of Pi by short time periods indicates that genes related to general stress are up-regulated before those specific to Pi signaling and metabolism. Our hydroponic system also proved useful for conducting pathogenesis essays, revealing early transcriptional activation of pathogenesis-related genes.

Transcriptomic data exploring the effect of agave fructans on the induction of the defense system in avocado fruit.

The effect of 20% high degree polymerized agave fructans (HDPAF) on the induction of the defense system in avocado fruits was investigated by transcriptomic analysis at 1, 24 and 72 h after treatment, and the effect of HDPAF on respiration rate and ethylene production was also analyzed. Transcriptomic profiling revealed 5425 differentially expressed genes (DEGs), 55 of which were involved in the pathways related to plant defense response to pathogens. Key genes were associated with phenylpropanoid biosynthesis, mitogen-activated protein signaling, plant hormone signaling, calcium ion signal decoding, and pathogenesis-related proteins. Dysregulated genes involved in ethylene biosynthesis were also identified, and the reduction in ethylene production by HDPAF was corroborated by gas chromatography, where three days of delayed peak production was observed compared to that in water-treated fruits. These results help to understand the mechanism of induction of the avocado defense system by applying HDPAF and support the application of HDPAF as an efficient postharvest treatment to extend the shelf life of the fruit.

Effect of Metabolites Produced by Bacillus atrophaeus and Brevibacterium frigoritolerans Strains on Postharvest Biocontrol of Alternaria alternata in Tomato (Solanum lycopersicum L.).

In the present study, the antifungal activity of metabolites produced by Bacillus atrophaeus B5 and a new Brevibacterium strain against Alternaria alternata was evaluated. Assays in vitro and in vivo on tomato fruit during postharvest were made. Based on the 16S rDNA gene sequence analysis, the new strain (strain B7) was identified as Brevibacterium frigoritolerans. Metabolites produced by both bacterial strains reduced the spore germination of A. alternata in vitro and decreased the severity of the alternaria rot disease on tomato fruit during postharvest. This is the first report that demonstrates the potential of B. frigoritolerans B7 as a biocontrol agent against this fungal phytopathogen. The use of metabolites produced by B. atrophaeus B5 and B. frigoritolerans B7 represents a new approach to reduce the use of chemical pesticides and control fungal decay during the postharvest stage.

The Pbo Cluster from Pseudomonas syringae pv. Phaseolicola NPS3121 Is Thermoregulated and Required for Phaseolotoxin Biosynthesis.

The bean (Phaseolus vulgaris) pathogen Pseudomonas syringae pv. phaseolicola NPS3121 synthesizes phaseolotoxin in a thermoregulated way, with optimum production at 18 °C. Gene PSPPH_4550 was previously shown to be thermoregulated and required for phaseolotoxin biosynthesis. Here, we established that PSPPH_4550 is part of a cluster of 16 genes, the Pbo cluster, included in a genomic island with a limited distribution in P. syringae and unrelated to the possession of the phaseolotoxin biosynthesis cluster. We identified typical non-ribosomal peptide synthetase, and polyketide synthetase domains in several of the pbo deduced products. RT-PCR and the analysis of polar mutants showed that the Pbo cluster is organized in four transcriptional units, including one monocistronic and three polycistronic. Operons pboA and pboO are both essential for phaseolotoxin biosynthesis, while pboK and pboJ only influence the amount of toxin produced. The three polycistronic units were transcribed at high levels at 18 °C but not at 28 °C, whereas gene pboJ was constitutively expressed. Together, our data suggest that the Pbo cluster synthesizes secondary metabolite(s), which could participate in the regulation of phaseolotoxin biosynthesis.

Activation of the phenylpropanoid biosynthesis pathway reveals a novel action mechanism of the elicitor effect of chitosan on avocado fruit epicarp.

Secondary metabolites play an important role in the avocado fruit defense system. Phenolic compounds are the main biosynthesized metabolites of this system response. Our objective in this investigation was to evaluate the induction of specific metabolic pathways using chitosan as an elicitor. Extracts obtained from avocado in intermediate and consumption maturity stages treated with chitosan exhibited an increase in antifungal activity, which caused inhibition of mycelial growth and a decrease in sporulation as well as spore germination of Colletotrichum gloeosporioides. Additionally, RNA from epicarp of the fruits treated and untreated with chitosan was obtained in order to evaluate the expression of genes related to phenylpropanoids and the antifungal compound 1-acetoxy-2-hydroxy-4-oxo-heneicosa-12,15-diene biosynthesis. An increased in gene expression of genes that participates in the phenylpropanoids route was observed during the stage of physiological fruit maturity, others genes such as Flavonol synthase (Fls), increased only in samples obtained from fruit treated with chitosan at consumption maturity. Our results reveal a new molecular mechanism where chitosan induces a specific accumulation of phenylpropanoids and antifungal diene; this partially explains avocado's resistance against fungal pathogens. Finally, we discuss the molecular connections between chitosan induction and gene expression to explain the biological events that orchestrate the resistance pathways in fruits.

Identification and characterization of a new Bacillus atrophaeus strain B5 as biocontrol agent of postharvest anthracnose disease in soursop (Annona muricata) and avocado (Persea americana).

Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous fruit, including soursop and avocado. The use of fungicides to maintain the high quality of fruit creates a potential health risk. One alternative to this problem is the biological control, which has been applied successfully during postharvest. The Bacillus species are one of the most studied biological agents against postharvest pathogens because accomplish their biocontrol performance by producing a variety of metabolites. In this study, we evaluated the activity of metabolites contained in the cell free supernatant, obtained from Bacillus strain B5 culture, against micelial growth and spore germination of two virulent strains of C. gloeosporioides isolated from soursop and avocado. On the basis of 16S rDNA gene sequence analysis, this strain was identified as Bacillus atrophaeus. A preventive treatment using cell free supernatant, reduced severity and incidence of anthracnose disease on harvested soursop and avocado fruit. B. atrophaeus strain B5 harbors genes involved in the production of antibiotics such as surfactin, bacillomycin and iturin, which could be contributing to the efficiency of the preventive treatment during postharvest. The antagonistic role of metabolites contained in the cell free supernatant against anthracnose disease, provide a new approach by which to attack this problem and can help reduce the use of chemical pesticides, environmental pollution, leading to the safer fruit preservation.

Enzyme activity and partial characterization of proteases obtained from Bromelia karatas fruit and compared with Bromelia pinguin proteases.

The enzymatic activity and partial characterization of proteases from Bromelia karatas fruits were evaluated and compared with Bromelia pinguin proteases. The specific activity increased twofold after partial purification in both proteases. Partially purified proteases from Bromelia karatas showed good specific activity at pH 6.0-8.0 and residual activity of 70-100% for 60 min at 37-60 °C, similar to Bromelia pinguin proteases. The K m value of proteases from Bromelia karatas was higher (253.32 µM) than that of Bromelia pinguin proteases (234.94 µM). The use of specific protease inhibitors indicated the presence of cysteine and serine proteases. Proteases with molecular weight of 66.2-97 and 21-31 kDa were detected. Bromelia karatas proteases registered 73% hydrolysis using a soy protein concentrate, similar to the enzyme activity of Bromelia pinguin proteases and commercial bromelain. These results demonstrate that Bromelia karatas proteases could be a potential alternative protease in the food industry.

Transcriptomic Analysis of Avocado Hass (Persea americana Mill) in the Interaction System Fruit-Chitosan-Colletotrichum.

Avocado (Persea americana) is one of the most important crops in Mexico as it is the main producer, consumer, and exporter of avocado fruit in the world. However, successful avocado commercialization is often reduced by large postharvest losses due to Colletotrichum sp., the causal agent of anthracnose. Chitosan is known to have a direct antifungal effect and acts also as an elicitor capable of stimulating a defense response in plants. However, there is little information regarding the genes that are either activated or repressed in fruits treated with chitosan. The aim of this study was to identify by RNA-seq the genes differentially regulated by the action of low molecular weight chitosan in the avocado-chitosan-Colletotrichum interaction system. The samples for RNA-seq were obtained from fruits treated with chitosan, fruits inoculated with Colletotrichum and fruits both treated with chitosan and inoculated with the fungus. Non-treated and non-inoculated fruits were also analyzed. Expression profiles showed that in short times, the fruit-chitosan system presented a greater number of differentially expressed genes, compared to the fruit-pathogen system. Gene Ontology analysis of differentially expressed genes showed a large number of metabolic processes regulated by chitosan, including those preventing the spread of Colletotrichum. It was also found that there is a high correlation between the expression of genes in silico and qPCR of several genes involved in different metabolic pathways.

Dietary patterns, nutritional profile, and body mass index in Mexican schoolchildren: a cross-sectional study

Childhood obesity is a serious public health problem in Mexico. In this study, childhoodspecific dietary patterns and their relationship with overweight-obesity prevalence, nutrient profiles and types of foods consumed were studied. A descriptive cross-sectional study of 724 randomly selected schoolchildren between 9 to 12 years old from Nayarit State, Mexico was performed. Data on anthropometric characteristics and food intake were recorded. Seven dietary patterns and three specific diets were identified by multivariate analysis. A dietary pattern characterized by high legume, snack and low beverage intake was negatively associated with weight and body mass index. The overall overweight and obesity prevalence was 20.2% and 20.6%, respectively. Diet type significantly influenced (p<0.05) protein, carbohydrates and fat intake but did not show correlation with the overweightobesity status. Simple sugars, candies, pastries and sweetened beverages appeared in all dietary patterns. Dietary patterns in countries with a wide gastronomic diversity should be considered to design preventive nutrition intervention programs(AU)

La obesidad infantil es un problema de salud pública en México. En este trabajo, se estudiaron los patrones dietéticos de escolares mexicanos y su relación con la prevalencia de sobrepeso-obesidad, el perfil de nutrientes y los principales grupos de alimentos consumidos. Se llevó a cabo un estudio transversal descriptivo donde participaron 724 escolares seleccionados al azar de entre 9 a 12 años del Estado de Nayarit, México evaluándose las características antropométricas y la ingesta de alimentos. Siete patrones dietéticos y tres dietas específicas fueron identificados mediante análisis multivariado. Un patrón de dieta caracterizada por una alta ingesta de legumbres y aperitivos junto con una baja ingesta de bebidas azucaradas se asoció negativamente con el peso e índice de masa corporal. La prevalencia de sobrepeso y obesidad fue del 20.2 y 20.6%, respectivamente. El tipo de dieta influyó significativamente (p<0.05) en la ingesta de proteínas, hidratos de carbono y el consumo de grasas, pero no mostró correlación con la condición de sobrepeso-obesidad. Los azúcares simples, dulces, postres y bebidas endulzadas aparecieron en todos los patrones dietéticos. Los patrones dietéticos en los países con una gran diversidad gastronómica podrían ser considerados de interés para diseñar programas de intervención nutricional preventiva(AU)

Global expression pattern comparison between low phosphorus insensitive 4 and WT Arabidopsis reveals an important role of reactive oxygen species and jasmonic acid in the root tip response to phosphate starvation.

Plants are exposed to several biotic and abiotic stresses. A common environmental stress that plants have to face both in natural and agricultural ecosystems that impacts both its growth and development is low phosphate (Pi) availability. There has been an important progress in the knowledge of the molecular mechanisms by which plants cope with Pi deficiency. However, the mechanisms that mediate alterations in the architecture of the Arabidopsis root system responses to Pi starvation are still largely unknown. One of the most conspicuous developmental effects of low Pi on the Arabidopsis root system is the inhibition of primary root growth that is accompanied by loss of root meristematic activity. To identify signalling pathways potentially involved in the Arabidpsis root meristem response to Pi-deprivation, here we report the global gene expression analysis of the root tip of wild type and low phosphorus insensitive4 (lpi4) mutant grown under Pi limiting conditions. Differential gene expression analysis and physiological experiments show that changes in the redox status, probably mediated by jasmonic acid and ethylene, play an important role in the primary root meristem exhaustion process triggered by Pi-starvation.

Characterization of low phosphorus insensitive mutants reveals a crosstalk between low phosphorus-induced determinate root development and the activation of genes involved in the adaptation of Arabidopsis to phosphorus deficiency.

Low phosphorus (P) availability is one of the most limiting factors for plant productivity in many natural and agricultural ecosystems. Plants display a wide range of adaptive responses to cope with low P stress, which generally serve to enhance P availability in the soil and to increase its uptake by roots. In Arabidopsis (Arabidopsis thaliana), primary root growth inhibition and increased lateral root formation have been reported to occur in response to P limitation. To gain knowledge of the genetic mechanisms that regulate root architectural responses to P availability, we designed a screen for identifying Arabidopsis mutants that fail to arrest primary root growth when grown under low P conditions. Eleven low phosphorus insensitive (lpi) mutants that define at least four different complementation groups involved in primary root growth responses to P availability were identified. The lpi mutants do not show the typical determinate developmental program induced by P stress in the primary root. Other root developmental aspects of the low P rescue system, including increased root hair elongation and anthocyanin accumulation, remained unaltered in lpi mutants. In addition to the insensitivity of primary root growth inhibition, when subjected to P deprivation, lpi mutants show a reduced induction in the expression of several genes involved in the P starvation rescue system (PHOSPHATE TRANSPORTER 1 and 2, PURPLE ACID PHOSPHATASE 1, ACID PHOSPHATASE 5, and INDUCED BY PHOSPHATE STARVATION 1). Our results provide genetic support for the role of P as an important signal for postembryonic root development and root meristem maintenance and show a crosstalk in developmental and biochemical responses to P deprivation.

Phosphate starvation induces a determinate developmental program in the roots of Arabidopsis thaliana.

When growing under limiting phosphate (P) conditions, Arabidopsis thaliana plants show dramatic changes in root architecture, including a reduction in primary root length, increased formation of lateral roots and greater formation of root hairs. Here we report that primary root growth inhibition by low P is caused by a shift from an indeterminate to a determinate developmental program. In the primary root, the low P-induced determinate growth program initiates with a reduction of cell elongation followed by the progressive loss of meristematic cells. At later stages, cell proliferation ceases and cell differentiation takes place at the former cell elongation and meristematic regions of the primary root. In low P, not only the primary but also almost all mature lateral roots enter the determinate developmental program. Kinetic studies of expression of the cell cycle marker CycB1;1:uidA and the quiescent center (QC) identity marker QC46:GUS showed that in low P conditions, reduction in proliferation in the primary root was preceded by alterations in the QC. These results suggest that in Arabidopsis, P limitation can induce a determinate root developmental program that plays an important role in altering root system architecture and that the QC could act as a sensor of environmental signals.