Challenges for Plant Growth Promoting Microorganism Transfer from Science to Industry: A Case Study from Chile.

Research on the plant growth promoting microorganisms (PGPM) is increasing strongly due to the biotechnological potential for the agricultural, forestry, and food industry. The benefits of using PGPM in crop production are well proven; however, their incorporation in agricultural management is still limited. Therefore, we wanted to explore the gaps and challenges for the transfer of biotechnological innovations based on PGPM to the agricultural sector. Our systematic review of the state of the art of PGPM research and knowledge transfer takes Chile as an example. Several transfer limiting aspects are identified and discussed. Our two main conclusions are: neither academia nor industry can meet unfounded expectations during technology transfer, but mutually clarifying their needs, capabilities, and limitations is the starting point for successful collaborations; the generation of a collaborative innovation environment, where academia as well as public and private stakeholders (including the local community) take part, is crucial to enhance the acceptance and integration of PGPM on the way to sustainable agriculture.

New Insights into Bacillus-Primed Plant Responses to a Necrotrophic Pathogen Derived from the Tomato-Botrytis Pathosystem.

Induced systemic resistance (ISR) is one of the most studied mechanisms of plant−microbe interaction and is considered a very promising alternative for integrated pest management programs. In our study, we explored the plant defense response induced by Bacillus velezensis BBC047 in relation to its application before or after Botrytis cinerea infection of tomato plants. The inoculation of BBC047 did not considerably alter the gene expression of the tomato tissues, whereas infection with B. cinerea in BBC047-primed plants induced expression of LRR and NBS-LRR receptors, which are highly related to the ISR response. As expected, B. cinerea infection generated molecular patterns typical of a defense response to pathogen infection as the overexpression of pathogenesis-related proteins (PRs) in leaflets distant to the point of infection. The curative treatment (P + F + B) allowed us to gain insights into plant response to an inverted priming. In this treatment, B. cinerea caused the m tissue damage, extending nearly entirely across the entire infected leaves. Additionally, genes generally associated with early SAR response (<16 h) were overexpressed, and apparently, the beneficial strain was not perceived as such. Therefore, we infer that the plant defense to the curative treatment represents a higher degree of biological stress triggered by the incorporation of strain BBC047 as second arriving microorganism. We highlight the importance the phytosanitary status of plants prior to inoculation of beneficial microorganism for the biocontrol of pathogens.

Metabolic imprint induced by seed halo-priming promotes a differential physiological performance in two contrasting quinoa ecotypes.

"Memory imprint" refers to the process when prior exposure to stress prepares the plant for subsequent stress episodes. Seed priming is a strategy to change the performance of seedlings to cope with stress; however, mechanisms associated with the metabolic response are fragmentary. Salinity is one of the major abiotic stresses that affect crop production in arid and semiarid areas. Chenopodium quinoa Willd. (Amaranthaceae) is a promising crop to sustain food security and possesses a wide genetic diversity of salinity tolerance. To elucidate if the metabolic memory induced by seed halo-priming (HP) differs among contrasting saline tolerance plants, seeds of two ecotypes of Quinoa (Socaire from Atacama Salar, and BO78 from Chilean Coastal/lowlands) were treated with a saline solution and then germinated and grown under different saline conditions. The seed HP showed a more positive impact on the sensitive ecotype during germination and promoted changes in the metabolomic profile in both ecotypes, including a reduction in carbohydrates (starch) and organic acids (citric and succinic acid), and an increase in antioxidants (ascorbic acid and α-tocopherol) and related metabolites. These changes were linked to a further reduced level of oxidative markers (methionine sulfoxide and malondialdehyde), allowing improvements in the energy use in photosystem II under saline conditions in the salt-sensitive ecotype. In view of these results, we conclude that seed HP prompts a "metabolic imprint" related to ROS scavenger at the thylakoid level, improving further the physiological performance of the most sensitive ecotype.

The Role of Surfactin Production by Bacillus velezensis on Colonization, Biofilm Formation on Tomato Root and Leaf Surfaces and Subsequent Protection (ISR) against Botrytis cinerea.

Many aspects regarding the role of lipopeptides (LPs) in bacterial interaction with plants are not clear yet. Of particular interest is the LP family of surfactin, immunogenic molecules involved in induced systemic resistance (ISR) and the bacterial colonization of plant surfaces. We hypothesize that the concentration of surfactin produced by a strain correlates directly with its ability to colonize and persist on different plant surfaces, which conditions its capacity to trigger ISR. We used two Bacillus velezensis strains (BBC023 and BBC047), whose antagonistic potential in vitro is practically identical, but not on plant surfaces. The surfactin production of BBC047 is 1/3 higher than that of BBC023. Population density and SEM images revealed stable biofilms of BBC047 on leaves and roots, activating ISR on both plant surfaces. Despite its lower surfactin production, strain BBC023 assembled stable biofilms on roots and activated ISR. However, on leaves only isolated, unstructured populations were observed, which could not activate ISR. Thus, the ability of a strain to effectively colonize a plant surface is not only determined through its production of surfactin. Multiple aspects, such as environmental stressors or compensation mechanisms may influence the process. Finally, the importance of surfactin lies in its impacts on biofilm formation and stable colonization, which finally enables its activity as an elicitor of ISR.

Importance of crop phenological stages for the efficient use of PGPR inoculants.

During the last decades, the incorporation of beneficial microorganisms in agriculture crop management has become a common practice. Seed coating of these microorganisms still faces technical issues, which limit its implementation in conventional agriculture. An adaption to widely established agricultural practices, e.g. fertigation, could help to overcome these issues. Here, using Bacillus velezensis strain BBC047, we show the influence of the crop phenological stages on the efficiency and success of microbial inoculation under agricultural conditions. In the commercial nursery, strain BBC047 improved growth in a variety of horticulture crops like basil, cabbage, tomato and bell pepper, the latter with the strongest effects in strengthening and accelerating the seedling growth (root and aerial biomass). For a field trial under productive conditions, different application strategies were compared, using bell pepper (Capsicum annuum L.) as crop under fertigation: conventional management (T1), application to the seedling (only nursery, T2), only post-transplant application (field, T3) and a combination of both (T4). In T2 and T4, the post-transplantation survival rate (p < 0.05) improved and the productivity of the plants increased (> 100%). Applications of BBC047 post-transplantation (T3) caused a lower increase in productivity (25%). Fruits from all three application strategies contained significantly more Vitamin C. We conclude that in conventional agriculture, the applications of PGPR inoculants to early crop phenological stages like nurseries are a viable alternative for the efficient use of PGPR inoculants. In comparison, a late introduction of a PGPR reduces its beneficial effect on crop productivity. We highlight that an appropriate timing in the use of PGPR inoculants is crucial for product development and success in sustainable agriculture.

Microbiome Dynamics Associated With the Atacama Flowering Desert.

In a desert, plants as holobionts quickly respond to resource pulses like precipitation. However, little is known on how environment and plants modulate the rhizosphere-associated microbiome. As a model species to represent the Atacama Desert bloom, Cistanthe longiscapa (Montiaceae family) was selected to study the influence of abiotic and biotic environment on the diversity and structure of the microbiota associated to its rhizosphere. We analyzed the rhizosphere and soil microbiome along a North-South precipitation gradient and between a dry and rainy year by using Illumina high-throughput sequencing of 16S rRNA gene fragments and ITS2 regions for prokaryotes and fungi, respectively. In the rhizosphere of C. longiscapa the microbiota clearly differs in composition and structure from the surrounding bulk soil. The fungal and bacterial communities respond differently to environmental conditions. The diversity and richness of fungal OTUs were negatively correlated with aridity, as predicted. The community structure was predominantly influenced by other soil characteristics (pH, organic matter content) but not by aridity. In contrast, diversity, composition, and structure of the bacterial community were not influenced by aridity or any other evaluated soil parameter. These findings coincide with the identification of mainly site-specific microbial communities, not shared along the sites. These local communities contain a group of OTUs, which are exclusive to the rhizosphere of each site and presumably vertically inherited as seed endophytes. Their ecological functions and dispersal mechanisms remain unclear. The analysis of co-occurrence patterns highlights the strong effect of the desert habitat over the soil- and rhizosphere-microbiome. The site-independent enrichment of only a small bacterial cluster consistently associated with the rhizosphere of C. longiscapa further supports this conclusion. In a rainy year, the rhizosphere microbiota significantly differed from bulk and bare soil, whereas in a dry year, the community structure of the former rhizosphere approximates to the one found in the bulk. In the context of plant-microbe interactions in desert environments, our study contributes new insights into the importance of aridity in microbial community structure and composition, discovering the influence of other soil parameters in this complex dynamic network, which needs further to be investigated.

Construction of a highly saturated linkage map in Japanese plum (Prunus salicina L.) using GBS for SNP marker calling.

This study reports the construction of high density linkage maps of Japanese plum (Prunus salicina Lindl.) using single nucleotide polymorphism markers (SNPs), obtained with a GBS strategy. The mapping population (An x Au) was obtained by crossing cv. "Angeleno" (An) as maternal line and cv. "Aurora" (Au) as the pollen donor. A total of 49,826 SNPs were identified using the peach genome V2.1 as a reference. Then a stringent filtering was carried out, which revealed 1,441 high quality SNPs in 137 An x Au offspring, which were mapped in eight linkage groups. Finally, the consensus map was built using 732 SNPs which spanned 617 cM with an average of 0.96 cM between adjacent markers. The majority of the SNPs were distributed in the intragenic region in all the linkage groups. Considering all linkage groups together, 85.6% of the SNPs were located in intragenic regions and only 14.4% were located in intergenic regions. The genetic linkage analysis was able to co-localize two to three SNPs over 37 putative orthologous genes in eight linkage groups in the Japanese plum map. These results indicate a high level of synteny and collinearity between Japanese plum and peach genomes.

De novo transcriptome assembly of 'Angeleno' and 'Lamoon' Japanese plum cultivars (Prunus salicina).

Japanese plum (Prunus salicina L.) is a fruit tree of the Rosaceae family, which is an economically important stone fruit around the world. Currently, Japanese plum breeding programs combine traditional breeding and plant physiology strategies with genetic and genomic analysis. In order to understand the flavonoid pathway regulation and to develop molecular markers associated to the fuit skin color (EST-SSRs), we performed a next generation sequencing based on Illumina Hiseq2000 platform. A total of 22.4 GB and 21 GB raw data were obtained from 'Lamoon' and 'Angeleno' respectively, corresponding to 85,404,726 raw reads to 'Lamoon' and 79,781,666 to 'Angeleno'. A total of 139,775,975 reads were filtered after removing low-quality reads and trimming the adapter sequences. De novo transcriptome assembly was performed using CLC Genome Workbench software and a total of 54,584 unique contigs were generated, with an N50 of 1343 base pair (bp) and a mean length of 829 bp. This work contributed with a specific Japanese plum skin transcriptome, providing two libraries of contrasting fruit skin color phenotype (yellow and red) and increasing substantially the GB of raw data available until now for this specie.

Genome-wide identification and characterization of R2R3MYB family in Rosaceae.

Transcription factors R2R3MYB family have been associated with the control of secondary metabolites, development of structures, cold tolerance and response to biotic and abiotic stress, among others. In recent years, genomes of Rosaceae botanical family are available. Although this information has been used to study the karyotype evolution of these species from an ancestral genome, there are no studies that treat the evolution and diversity of gene families present in these species or in the botanical family. Here we present the first comparative study of the R2R3MYB subfamily of transcription factors in three species of Rosaceae family (Malus domestica, Prunus persica and Fragaria vesca). We described 186, 98 and 86 non-redundant gene models for apple, peach and strawberry, respectively. In this research, we analyzed the intron-exon structure and genomic distribution of R2R3MYB families mentioned above. The phylogenetic comparisons revealed putative functions of some R2R3MYB transcription factors. This analysis found 44 functional subgroups, seven of which were unique for Rosaceae. In addition, our results showed a highly collinearity among some genes revealing the existence of conserved gene models between the three species studied. Although some gene models in these species have been validated under several approaches, more research in the Rosaceae family is necessary to determine gene expression patterns in specific tissues and development stages to facilitate understanding of the regulatory and biochemical mechanism in this botanical family.

Analysis of anthocyanin biosynthesis genes expression profiles in contrasting cultivars of Japanese plum (Prunus salicina L.) during fruit development.

Flavonoids are responsible of different fruit sensorial properties. In Japanese plum (Prunus salicina L.) these compounds are variable in both type and quantity during the different stages of fruit growth and maturation. Here we present the first study which determines the expression profile of structural genes of the flavonoid pathway and accumulation profiles of total phenols, proanthocyanidins and anthocyanins during fruit development stages in contrasting cultivars in Japanese plum. The biosynthesis of these compounds is differentially regulated in different tissues and cultivars. Our result showed that all pigmented tissues increased the expression of the leucoanthocyanidin dioxygenase (LDOX) gene, while all tissues without anthocyanin accumulation presented a minimal expression of LDOX. In addition, the regulation of putative transcription factors PsMYB10 and PsMYB1 were correlated positively and negatively with the pigmented tissues respectively, suggesting a critical and coordinated mechanism involved in the change of the fruit color.

Overexpression of GlyI and GlyII genes in transgenic tomato (Solanum lycopersicum Mill.) plants confers salt tolerance by decreasing oxidative stress.

The glyoxalase system plays an important role in various physiological processes in plants, including salt stress tolerance. We report the effects of overexpressing glyoxalase I and glyoxalase II genes in transgenic tomato (Solanum lycopersicum Mill.) cv. Ailsa Craig. Stable expression of both transgenes was detected in the transformed tomato plants under salt stress. The transgenic lines overexpressing GlyI and GlyII under a high NaCl concentration (800 mM) showed reduced lipid peroxidation and the production of H2O2 in leaf tissues. A greater decrease in the chlorophyll a+b content in wild-type (WT) compared with transgenic lines was also observed. These results suggest that the over expression of two genes, GlyI and GlyII, may enhance salt stress tolerance by decreasing oxidative stress in transformed tomato plants. This work will help our understanding of the putative role of the glyoxalase system in the tolerance to abiotic stress in tomato plants.

Bilateral simultaneous ectopic pregnancy.

Spontaneous bilateral ectopic pregnancy is the rarest form of ectopic pregnancy. A 31-year-old patient with spontaneous bilateral tubal pregnancy was admitted after light vaginal bleeding, 42 days after her last menstruation. Ultrasonography showed a bilateral tubal ectopic pregnancy. A bilateral laparoscopic salpingectomy was performed without complication and the pathology report confirmed the diagnosis. This is an unusual case of early diagnosis.

Aspectos clínicos más relevantes de la cirugía de los terceros molares incluidos

Se ha demostrado que el 17 porciento de la poblaci¢n actual mayor de 20 años, presenta uno o más dientes retenidos. El procedimiento quirúrgico que más se realiza en los servicios de Cirugía Maxilo Facial es la extracción de los terceros molares incluidos. Se realiza un estudio con el propósito de describir aspectos clínicos más relevantes de la cirugía de los terceros molares en el servicio de cirugía maxilo facial del Hospital Vladimir I. Lenin de Holguín durante el año 2000. Se intervienen quirúgicamente setenta y seis pacientes que constituyó el universo de trabajo, en los resultados la mayor frecuencia de remisiones estuvo entre los 18 y 24 años de edad, la causa de mayor número de remisiones fue la periconaritis con un 65,7 por ciento. El grado de retenci¢n más encontrada fue la submucosa con un 65,7 por ciento el edema fue la complicación postoperatoria más frecuente con un 97 por ciento seguida del trismo(AU)

Eventos estresantes en alumnos de cine y televisión / Stressful events in TV-film students

Después de caracterizar la Escuela Internacional de Cine y Televisión (San Antonio de los Baños, Habana, Cuba), y a los alumnos que en ella se forman, se relacionan seis grupos de eventos desencadenantes de estrés, que según los resultados obtenidos mediante la exploración directa a 50 estudiantes de dicho centro, son los que, en mayor medida, provocan distrés, y consecuentemente, la aparición de afecciones psicosomáticas. Se describen, además, cuáles son las reacciones más frecuentes, así como las medidas terapéuticas aplicadas con regularidad

Eventos estresantes en alumnos de cine y televisión / Stressful events in TV-film students

Después de caracterizar la Escuela Internacional de Cine y Televisión (San Antonio de los Baños, Habana, Cuba), y a los alumnos que en ella se forman, se relacionan seis grupos de eventos desencadenantes de estrés, que según los resultados obtenidos mediante la exploración directa a 50 estudiantes de dicho centro, son los que, en mayor medida, provocan distrés, y consecuentemente, la aparición de afecciones psicosomáticas. Se describen, además, cuáles son las reacciones más frecuentes, así como las medidas terapéuticas aplicadas con regularidad(AU)