The roles of WRKY transcription factors in Malus spp. and Pyrus spp.

The WRKY transcription factor gene family is known to be involved in plant defense against pathogens and in tolerance to different environmental stresses at different stages of development. The response mechanisms through which these genes act can be influenced by different phytohormones as well as by many trans- and cis-acting elements, making this network an important topic for analysis, but still something complex to fully understand. According to available reports, these genes can also perform important roles in pome species (Malus spp. and Pyrus spp.) metabolism, especially in adaptation of these plants to stressful conditions. Here, we present a quick review of what is known about WRKY genes in Malus and Pyrus genomes offering a simple way to understand what is already known about this topic. We also add information connecting the evolution of these transcription factors with others that can also be found in pomes.

Morphological, sensory and biochemical characteristics of summer apple genotypes / Características morfológicas, sensoriais e bioquímicas de genótipos de maçã cultivada no verão

Summer apples are one of the most important plant community in Artvin province located Northeastern part of Turkey. In present study 22 local apple genotypes were characterized by phenological, morphological, biochemical and sensory properties. Harvest date was the main phenological data. Morphological measurements included fruit weight, fruit shape, fruit ground color, fruit over color, fruit over color coverage and fruit firmness, respectively. Sensory measurements were as juiciness and aroma and biochemical characteristics included organic acids, SSC (Soluble Solid Content), vitamin C, total phenolic content and antioxidant capacity. Genotypes exhibited variable harvest dates ranging from 11 July to 13 August and cv. Summered harvested 30 July 2017. The majority of genotypes were harvested before cv. Summered. Fruit weight were also quite variable among genotypes which found to be between 89 g and 132 g, and most of the genotypes had bigger fruits than cv. Summered. Pink, red, yellow and green fruit skin color was evident and main fruit shape were determined as round, conic and oblate among genotypes. ART08-9, ART08-4, ART08-21 and ART08-22 had distinct bigger fruits and ART08-1, ART08-2, ART08-5, ART08-12 and ART08-17 had higher total phenolic content and antioxidant capacity. The results of the study showed significant differences for most of the phenological, morphological, sensory and biochemical characteristics. Thus, the phonological, morphological, sensory and biochemical characteristics of summer apple genotypes were distinguishable and these results suggest that phonological, morphological, sensory and biochemical differences of the summer apple genotypes can be attributed to differences in genetic background of genotypes which placed different groups by PCoA analysis.(AU)

As maçãs cultivadas no verão são uma das culturas vegetais mais importantes da província de Artvin, localizada no nordeste da Turquia. No presente estudo, 22 genótipos locais de maçã foram caracterizados quanto às suas propriedades morfológicas, bioquímicas e sensoriais. As características analisadas foram peso do fruto, data da colheita, forma do fruto, coloração da casca, firmeza do fruto, suculência, aroma, teor de ácidos orgânicos e de sólidos solúveis, vitamina C, teor fenólico total e capacidade antioxidante. Os genótipos exibiram datas de colheita que variaram de 11 de julho a 13 de agosto, e a cultivar de verão foi colhida em 30 de julho 2017. A maioria dos genótipos foi colhida antes da cultivar de verão. O peso dos frutos também foi bastante variável entre os genótipos, apresentando entre 89 e 132 g, e a maioria dos genótipos apresentou frutos maiores que a cultivar de verão. As colorações rosa, vermelho, amarelo e verde da casca dos frutos foram as mais evidentes, e a forma principal dos frutos foi determinada como redonda, cônica e oblata entre os genótipos. ART08-9, ART08-4, ART08-21 e ART08-22apresentaram frutos maiores distintos, e ART08-1, ART08-2, ART08-5, ART08-12 e ART08-17 apresentaram maior conteúdo fenólico total e capacidade antioxidante. Os resultados do estudo mostraram diferenças significativas para a maioria das características morfológicas, sensoriais e bioquímicas. Assim, essas características dos genótipos da maçã cultivadas no verão foram distintos, e esses resultados podem ser atribuídos a diferenças no contexto genético dos genótipos.(AU)

Horticultural performance of 'Fuji Suprema' and 'Maxi Gala' apple trees trained at Tall Spindle and Bi-Axis systems grafted on different rootstocks in Southern Brazil / Comportamento agronômico de macieiras 'Fuji Suprema' e 'Maxi Gala' conduzidas em Tall Spindle e Bi-Axis enxertadas em diferentes porta-enxertos no Sul do Brasil

The use of new rootstocks combined with different training systems have been studied to increase productivity, fruit quality, and reduce costs. Another important factor is the validation of studies under replanting conditions, due to the growing limitation of new areas for planting commercial apple orchards in Brazil. The present study aimed to evaluate the behavior of 'Fuji Suprema' and 'Maxi Gala' apple trees in two training systems grafted on two rootstocks. The study occurred during the seasons 2016/17, 2017/18 and 2018/19, in an orchard located in southern Brazil in an area previously cultivated with apple trees. A randomized block design consisting of a 2x2 factorial was used, with Tall Spindle and Bi-Axis as the training system and 'M.9' and 'G.213' as rootstocks of Fuji Suprema and Maxi Gala cultivars. Variables were evaluated to determine tree vigor, as well as yield and fruit quality characteristics. It was observed that 'G.213' promoted higher vigor for both cultivars as well as the Bi-Axis training system in the Maxi Gala cultivar. The highest yield was observed in Tall Spindle as well as the rootstock 'G.213' in both cultivars. The most significant changes on fruit quality occurred in the Bi-Axis training system, presenting higher intensity of red color as well as 'Maxi Gala' grafted on 'G.213' rootstock.

A utilização de novos porta-enxertos e diferentes sistemas de condução tem sido estudado para aumentar a produtividade, qualidade dos frutos e reduzir custos. Outro fator importante é a validação de estudos em condições de replantio, devido à crescente limitação de novas áreas para o plantio comercial de pomares de macieiras no Brasil. O presente estudo teve como objetivo avaliar o comportamento de macieiras 'Fuji Suprema' e 'Maxi Gala' em dois sistemas de condução enxertados sobre dois porta-enxertos. O estudo ocorreu durante as safras 2016/17, 2017/18 e 2018/19, em um pomar localizado no sul do Brasil, em área previamente cultivada com macieiras. Utilizou-se delineamento de blocos casualizados, consistindo em um fatorial 2x2, sendo Tall Spindle e Bi-Axis como sistemas de condução e 'M.9' e 'G.213' como porta-enxertos das cultivares Fuji Suprema e Maxi Gala. Foram avaliadas variáveis para determinar o comportamento vegetativo, bem como as características de produtividade e qualidade dos frutos. Observou-se que 'G.213' promoveu maior vigor para ambas cultivares, assim como o sistema de condução Bi-Axis na cultivar Maxi Gala. O maior rendimento foi observado nas macieiras conduzidas em Tall Spindle e enxertadas sobre 'G.213' em ambas cultivares. As mudanças mais significativas na qualidade dos frutos ocorreram nas macieiras conduzidas em Bi-Axis, apresentando maior intensidade de coloração vermelha, assim como na macieira 'Maxi Gala' enxertada sobre 'G.213'.

Morphological, sensory and biochemical characteristics of summer apple genotypes / Características morfológicas, sensoriais e bioquímicas de genótipos de maçã cultivada no verão

Summer apples are one of the most important plant community in Artvin province located Northeastern part of Turkey. In present study 22 local apple genotypes were characterized by phenological, morphological, biochemical and sensory properties. Harvest date was the main phenological data. Morphological measurements included fruit weight, fruit shape, fruit ground color, fruit over color, fruit over color coverage and fruit firmness, respectively. Sensory measurements were as juiciness and aroma and biochemical characteristics included organic acids, SSC (Soluble Solid Content), vitamin C, total phenolic content and antioxidant capacity. Genotypes exhibited variable harvest dates ranging from 11 July to 13 August and cv. Summered harvested 30 July 2017. The majority of genotypes were harvested before cv. Summered. Fruit weight were also quite variable among genotypes which found to be between 89 g and 132 g, and most of the genotypes had bigger fruits than cv. Summered. Pink, red, yellow and green fruit skin color was evident and main fruit shape were determined as round, conic and oblate among genotypes. ART08-9, ART08-4, ART08-21 and ART08-22 had distinct bigger fruits and ART08-1, ART08-2, ART08-5, ART08-12 and ART08-17 had higher total phenolic content and antioxidant capacity. The results of the study showed significant differences for most of the phenological, morphological, sensory and biochemical characteristics. Thus, the phonological, morphological, sensory and biochemical characteristics of summer apple genotypes were distinguishable and these results suggest that phonological, morphological, sensory and biochemical differences of the summer apple genotypes can be attributed to differences in genetic background of genotypes which placed different groups by PCoA analysis.

As maçãs cultivadas no verão são uma das culturas vegetais mais importantes da província de Artvin, localizada no nordeste da Turquia. No presente estudo, 22 genótipos locais de maçã foram caracterizados quanto às suas propriedades morfológicas, bioquímicas e sensoriais. As características analisadas foram peso do fruto, data da colheita, forma do fruto, coloração da casca, firmeza do fruto, suculência, aroma, teor de ácidos orgânicos e de sólidos solúveis, vitamina C, teor fenólico total e capacidade antioxidante. Os genótipos exibiram datas de colheita que variaram de 11 de julho a 13 de agosto, e a cultivar de verão foi colhida em 30 de julho 2017. A maioria dos genótipos foi colhida antes da cultivar de verão. O peso dos frutos também foi bastante variável entre os genótipos, apresentando entre 89 e 132 g, e a maioria dos genótipos apresentou frutos maiores que a cultivar de verão. As colorações rosa, vermelho, amarelo e verde da casca dos frutos foram as mais evidentes, e a forma principal dos frutos foi determinada como redonda, cônica e oblata entre os genótipos. ART08-9, ART08-4, ART08-21 e ART08-22apresentaram frutos maiores distintos, e ART08-1, ART08-2, ART08-5, ART08-12 e ART08-17 apresentaram maior conteúdo fenólico total e capacidade antioxidante. Os resultados do estudo mostraram diferenças significativas para a maioria das características morfológicas, sensoriais e bioquímicas. Assim, essas características dos genótipos da maçã cultivadas no verão foram distintos, e esses resultados podem ser atribuídos a diferenças no contexto genético dos genótipos.

Morphological, sensory and biochemical characteristics of summer apple genotypes / Características morfológicas, sensoriais e bioquímicas de genótipos de maçã cultivada no verão

Summer apples are one of the most important plant community in Artvin province located Northeastern part of Turkey. In present study 22 local apple genotypes were characterized by phenological, morphological, biochemical and sensory properties. Harvest date was the main phenological data. Morphological measurements included fruit weight, fruit shape, fruit ground color, fruit over color, fruit over color coverage and fruit firmness, respectively. Sensory measurements were as juiciness and aroma and biochemical characteristics included organic acids, SSC (Soluble Solid Content), vitamin C, total phenolic content and antioxidant capacity. Genotypes exhibited variable harvest dates ranging from 11 July to 13 August and cv. Summered harvested 30 July 2017. The majority of genotypes were harvested before cv. Summered. Fruit weight were also quite variable among genotypes which found to be between 89 g and 132 g, and most of the genotypes had bigger fruits than cv. Summered. Pink, red, yellow and green fruit skin color was evident and main fruit shape were determined as round, conic and oblate among genotypes. ART08-9, ART08-4, ART08-21 and ART08-22 had distinct bigger fruits and ART08-1, ART08-2, ART08-5, ART08-12 and ART08-17 had higher total phenolic content and antioxidant capacity. The results of the study showed significant differences for most of the phenological, morphological, sensory and biochemical characteristics. Thus, the phonological, morphological, sensory and biochemical characteristics of summer apple genotypes were distinguishable and these results suggest that phonological, morphological, sensory and biochemical differences of the summer apple genotypes can be attributed to differences in genetic background of genotypes which placed different groups by PCoA analysis.

As maçãs cultivadas no verão são uma das culturas vegetais mais importantes da província de Artvin, localizada no nordeste da Turquia. No presente estudo, 22 genótipos locais de maçã foram caracterizados quanto às suas propriedades morfológicas, bioquímicas e sensoriais. As características analisadas foram peso do fruto, data da colheita, forma do fruto, coloração da casca, firmeza do fruto, suculência, aroma, teor de ácidos orgânicos e de sólidos solúveis, vitamina C, teor fenólico total e capacidade antioxidante. Os genótipos exibiram datas de colheita que variaram de 11 de julho a 13 de agosto, e a cultivar de verão foi colhida em 30 de julho 2017. A maioria dos genótipos foi colhida antes da cultivar de verão. O peso dos frutos também foi bastante variável entre os genótipos, apresentando entre 89 e 132 g, e a maioria dos genótipos apresentou frutos maiores que a cultivar de verão. As colorações rosa, vermelho, amarelo e verde da casca dos frutos foram as mais evidentes, e a forma principal dos frutos foi determinada como redonda, cônica e oblata entre os genótipos. ART08-9, ART08-4, ART08-21 e ART08-22 apresentaram frutos maiores distintos, e ART08-1, ART08-2, ART08-5, ART08-12 e ART08-17 apresentaram maior conteúdo fenólico total e capacidade antioxidante. Os resultados do estudo mostraram diferenças significativas para a maioria das características morfológicas, sensoriais e bioquímicas. Assim, essas características dos genótipos da maçã cultivadas no verão foram distintos, e esses resultados podem ser atribuídos a diferenças no contexto genético dos genótipos.

Morphological, sensory and biochemical characteristics of summer apple genotypes.

Summer apples are one of the most important plant community in Artvin province located Northeastern part of Turkey. In present study 22 local apple genotypes were characterized by phenological, morphological, biochemical and sensory properties. Harvest date was the main phenological data. Morphological measurements included fruit weight, fruit shape, fruit ground color, fruit over color, fruit over color coverage and fruit firmness, respectively. Sensory measurements were as juiciness and aroma and biochemical characteristics included organic acids, SSC (Soluble Solid Content), vitamin C, total phenolic content and antioxidant capacity. Genotypes exhibited variable harvest dates ranging from 11 July to 13 August and cv. Summered harvested 30 July 2017. The majority of genotypes were harvested before cv. Summered. Fruit weight were also quite variable among genotypes which found to be between 89 g and 132 g, and most of the genotypes had bigger fruits than cv. Summered. Pink, red, yellow and green fruit skin color was evident and main fruit shape were determined as round, conic and oblate among genotypes. ART08-9, ART08-4, ART08-21 and ART08-22 had distinct bigger fruits and ART08-1, ART08-2, ART08-5, ART08-12 and ART08-17 had higher total phenolic content and antioxidant capacity. The results of the study showed significant differences for most of the phenological, morphological, sensory and biochemical characteristics. Thus, the phonological, morphological, sensory and biochemical characteristics of summer apple genotypes were distinguishable and these results suggest that phonological, morphological, sensory and biochemical differences of the summer apple genotypes can be attributed to differences in genetic background of genotypes which placed different groups by PCoA analysis.

Functional characterisation and in silico modelling of MdPSY2 variants and MdPSY5 phytoene synthases from Malus domestica.

Carotenoids are plastid isoprenoid pigments that play critical roles in light harvesting, photoprotection, and phytohormone biosynthesis. They are also vitamin-A precursors and antioxidant molecules important for human nutrition. Apples (e.g. Malus x domestica Borkh), one of the most widely consumed fruits with high nutrient levels, have a very low carotenoid concentration in flesh, compared with other fruits and vegetables. This could be explained by a deficiency in carotenoid synthesis/accumulation and/or accelerated degradation. We analysed the contribution of M. domestica cv. 'Fuji' phytoene synthase (PSY) in the biosynthesis of carotenoids and determined that among four MdPSY genes present in the organism, MdPSY2 and MdPSY5 are highly expressed in leaves and during fruit ripening in line with an increment in carotenoid content in fruits. Furthermore, two representative polymorphic MdPSY2 variants were found, one with a Tyr358Phe substitution (MdPSY2_F) and the other that additionally has a six-amino-acid deletion in the signal peptide (MdPSY2_CG). MdPSY2, MdPSY5, MdPSY2_F and MdPSY2_CG are all localised in plastids. Interestingly, the polymorphic MdPSY2_F and MdPSY2_CG variants show lower enzymatic activity than the wild-type form in a heterologous complementation assay, which could be attributed to the Tyr358Phe substitution close to the active-site pocket, as was suggested by 3-D modelling analysis. The presence of polymorphic MdPSY2 variants with lower enzymatic activity could be partially responsible for the low carotenoid content in Fuji apple fruits.

Boosting carotenoid content in Malus domestica var. Fuji by expressing AtDXR through an Agrobacterium-mediated transformation method.

Apple (Malus domestica) fruits accumulate negligible levels of carotenoids, antioxidant pigments that are precursors for vitamin A in humans. As vitamin A deficiency is an important public health issue, we aimed at increasing carotenoids in apple by constitutively expressing the Arabidopsis thaliana DXR gene, one of the key regulatory steps in the plastidial isoprenoid pathway. For this purpose, we optimized an Agrobacterium-mediated transformation method in the commercial Fuji Raku Raku variety. This resulted in a shoot establishment efficiency of 0.75% at 20 weeks after infection. Molecular and microscopical analyses revealed that 80% of the hygromycin resistant shoots contained and expressed AtDXR:eGFP and that the AtDXR:eGFP fusion protein located in plastids. Transgenic seedlings displayed up to 3-fold increase in total carotenoids and in individual carotenoids compared to the WT, correlating with an increased transcript abundance of endogenous carotenogenic genes such as MdDXS, MdPSY1, MdPSY2, MdPSY3, MdLCYB1, and MdLCYB2. In addition, buds of 2-year-old transgenic dormant trees showed an increment up to 3-fold in lutein, and transient transformation of fruits revealed that AtDXR induced a 2-fold increment in total carotenoids. Thus, these results suggest that DXR may be a good candidate for increasing carotenoid levels in apple fruits through metabolic engineering.

Serological and Molecular Detection of Latent Viruses in the Apple Germplasm Bank of Santa Catarina

Abstract The Apple Germplasm Bank (AGB) of Santa Catarina Agricultural Research and Rural Extension Company - Epagri, AGB-Epagri, is the largest of the genus Malus in Brazil. Twenty-eight main accessions of this bank were virus screened through DAS-ELISA, RT-PCR and IC-RT-PCR during two consecutive reproductive cycles, and each accession showed latent mixed infection by at least two species, among ASGV, ASPV and ACLSV. The combined use of diagnostic methods helped overcome inconsistencies commonly found in apple virus detection and was shown essential for the AGB-Epagri can be safely used as a source of genetic variability and for the exchange of virus-free propagative material.

Characterization of the nucellus-specific dehydrin MdoDHN11 demonstrates its involvement in the tolerance to water deficit.

KEY MESSAGE: MdoDHN11 acts in the nucellus layer to protect the embryo and the endosperm from limited water availability during apple seed development. Dehydrins (DHNs) are protective proteins related to several plant developmental responses that involve dehydration such as seed desiccation and abiotic stresses. In apple (Malus × domestica Borkh.), the seed-specific MdoDHN11 was suggested to play important roles against dehydration during seed development. However, this hypothesis has not yet been evaluated. Within this context, several experiments were performed to functionally characterize MdoDHN11. In situ hybridization analysis during apple seed development showed that MdoDHN11 expression is confined to a maternal tissue called nucellus, a central mass of parenchyma between the endosperm and the testa. The MdoDHN11 protein was localized in the cytosol and nucleus. Finally, transgenic Arabidopsis plants expressing MdoDHN11 were generated and exposed to a severe water-deficit stress, aiming to mimic a situation that can occurs during seed development. All transgenic lines showed increased tolerance to water deficit in relation to wild-type plants. Taken together, our results provide evidences that MdoDHN11 plays important roles during apple seed development by protecting the embryo and the endosperm from limited water availability, and the mechanism of action probably involves the interaction of MdoDHN11 with proteins and other components in the cell.

Phenotypic and molecular characterization of apple (Malus × domestica Borkh) genetic resources in Greece

The phenotypic and genetic analysis of apple (Malus × domestica Borkh) genotypes is essential for breeding species. Information on the morphology and genetic structure of apple offers significant help for germplasm maintenance and selection of suitable material to breed superior cultivars. This study shows the results of an investigation on the morphology and the genetic diversity for 19 apple cultivars, which are preserved in an ex situ collection in Naoussa, Central Macedonia, Greece. Information was recorded over a 5-year period for 47 traits describing plant morphology and phenotype, as well as leaf and fruit quality. Data were analyzed using the principal component analysis (PCA) method. The Euclidean distance metric and the Wards agglomeration method were used in an unsupervised hierarchical cluster analysis of all cultivars. The cultivars were grouped into four main clusters, suggesting that the characterized apple collection has a high potential for specific breeding goals. Furthermore, the cultivars were genotyped using seven microsatellite primers. Moderate levels of polymorphism were detected, and 38 distinctive alleles (5.4 alleles per primer pair) were identified. Both multivariate clustering approach (phenotypic data) and the genetic distance clustering approach (genetic data) grouped the apple cultivars according to their type. Hence, these data could be used for protection or patenting processes of existing or new apple cultivars carried out by the EU-Community Plant Variety Office.(AU)

Evolutionary diversification of galactinol synthases in Rosaceae: adaptive roles of galactinol and raffinose during apple bud dormancy.

Galactinol synthase (GolS) is a key enzyme in the biosynthetic pathway of raffinose family oligosaccharides (RFOs), which play roles in carbon storage, signal transduction, and osmoprotection. The present work assessed the evolutionary history of GolS genes across the Rosaceae using several bioinformatic tools. Apple (Malus × domestica) GolS genes were transcriptionally characterized during bud dormancy, in parallel with galactinol and raffinose measurements. Additionally, MdGolS2, a candidate to regulate seasonal galactinol and RFO content during apple bud dormancy, was functionally characterized in Arabidopsis. Evolutionary analyses revealed that whole genome duplications have driven GolS gene evolution and diversification in Rosaceae speciation. The strong purifying selection identified in duplicated GolS genes suggests that differential gene expression might define gene function better than protein structure. Interestingly, MdGolS2 was differentially expressed during bud dormancy, concomitantly with the highest galactinol and raffinose levels. One of the intrinsic adaptive features of bud dormancy is limited availability of free water; therefore, we generated transgenic Arabidopsis plants expressing MdGolS2. They showed higher galactinol and raffinose contents and increased tolerance to water deficit. Our results suggest that MdGolS2 is the major GolS responsible for RFO accumulation during apple dormancy, and these carbohydrates help to protect dormant buds against limited water supply.

Contrasting Transcriptional Programs Control Postharvest Development of Apples (Malus x domestica Borkh.) Submitted to Cold Storage and Ethylene Blockage.

Apple is commercially important worldwide. Favorable genomic contexts and postharvest technologies allow year-round availability. Although ripening is considered a unidirectional developmental process toward senescence, storage at low temperatures, alone or in combination with ethylene blockage, is effective in preserving apple properties. Quality traits and genome wide expression were integrated to investigate the mechanisms underlying postharvest changes. Development and conservation techniques were responsible for transcriptional reprogramming and distinct programs associated with quality traits. A large portion of the differentially regulated genes constitutes a program involved in ripening and senescence, whereas a smaller module consists of genes associated with reestablishment and maintenance of juvenile traits after harvest. Ethylene inhibition was associated with a reversal of ripening by transcriptional induction of anabolic pathways. Our results demonstrate that the blockage of ethylene perception and signaling leads to upregulation of genes in anabolic pathways. We also associated complex phenotypes to subsets of differentially regulated genes.

Modulation of oxidative responses by a virulent isolate of Colletotrichum fructicola in apple leaves.

Apple bitter rot (ABR) and Glomerella leaf spot (GLS) can be caused by Colletotrichum fructicola. Although both diseases can occur simultaneously in orchards, some isolates show clear organ specialization. Thus, this work was aimed to compare microscopically the development of preinfective structures of ABR- and GLS isolates and their impact on the enzymatic oxidant defense system during the leaf infection process. On leaves, conidial germlings of GLS-isolate formed appressoria mostly sessile. In contrast, those of ABR-isolate were pedicellate and formed multiple melanized appressoria probably as a sign of unsuccessful infection attempts. Neither ABR- nor GLS isolate triggered hypersensitive response in apple leaves. In overall, the activity of scavenging enzymes was higher and long-lasting in leaves inoculated by GLS- than by ABR isolate and control. Guaiacol peroxidase, catalase, and glutathione reductase had activity peaks within 24 h after inoculation (HAI). Ascorbate peroxidase activity was higher only in GLS-infected leaves at 6 HAI, while superoxide dismutase remained unaltered. A lower level of hydrogen peroxide (H2O2) was determined in GLS-infected plants at 48 HAI, but the electrolyte leakage markedly increased. Disease symptoms in leaves were only caused by GLS-isolate. Results suggest that the virulent isolate coordinately downregulates the oxidative plant defense responses enabling its successful establishment in apple leaves.

Analysis of 'Fuji' apple somatic variants from next-generation sequencing.

The domesticated apple (Malus x domestica Borkh.) is a major fruit crop of temperate regions of the world. 'Fuji' apple (Ralls Genet x Delicious), a famous apple cultivar in Korea, has been very popular since its promotion in Japan in 1958. 'Fuji' and its bud mutant cultivars possess variable levels of genetic diversity. Nonetheless, the phenotypes of each group, which are classified into the bud mutation groups: early season, fruiting spur, and coloring, are similar. Despite attempts to identify these bud mutation cultivars, molecular markers, which were developed before the emergence of next-generation sequencing technology, have not been able to distinguish each cultivar easily. In this study, we adopted the resequencing technique using the 'Golden Delicious' (Grimes Golden x Unknown) apple genome as a reference. SNPs (single nucleotide polymorphisms) and InDels (insertions or deletions) of 'Fuji' apple and its bud mutant cultivar were detected and SNPs and unique InDels distinct to each cultivar were identified. Data from this study may be used to identify bud mutant cultivars of 'Fuji' apples and be useful for further breeding of apples.

Cloning and characterization of the SERK1 gene in triploid Pingyi Tiancha [Malus hupehensis (Pamp.) Rehd. var. pingyiensis Jiang] and a tetraploid hybrid strain.

This study aims to explore the roles of somatic embryogenesis receptor-like kinase (SERK) in Malus hupehensis (Pingyi Tiancha). The full-length sequences of SERK1 in triploid Pingyi Tiancha (3n) and a tetraploid hybrid strain 33# (4n) were cloned, sequenced, and designated as MhSERK1 and MhdSERK1, respectively. Multiple alignments of amino acid sequences were conducted to identify similarity between MhSERK1 and MhdSERK1 and SERK sequences in other species, and a neighbor-joining phylogenetic tree was constructed to elucidate their phylogenetic relations. Expression levels of MhSERK1 and MhdSERK1 in different tissues and developmental stages were investigated using quantitative real-time PCR. The coding sequence lengths of MhSERK1 and MhdSERK1 were 1899 bp (encoding 632 amino acids) and 1881 bp (encoding 626 amino acids), respectively. Sequence analysis demonstrated that MhSERK1 and MhdSERK1 display high similarity to SERKs in other species, with a conserved intron/exon structure that is unique to members of the SERK family. Additionally, the phylogenetic tree showed that MhSERK1 and MhdSERK1 clustered with orange CitSERK (93%). Furthermore, MhSERK1 and MhdSERK1 were mainly expressed in the reproductive organs, in particular the ovary. Their expression levels were highest in young flowers and they differed among different tissues and organs. Our results suggest that MhSERK1 and MhdSERK1 are related to plant reproduction, and that MhSERK1 is related to apomixis in triploid Pingyi Tiancha.

Características de las residencias de terapia intensiva pediátrica de la República Argentina: Encuesta nacional / Characteristics of pediatric intensive care residency programs in Argentina: A national survey

Introducción. La residencia de terapia intensiva pediátrica (TIP) tiene pocos años de desarrollo en nuestro país. Conocer su situación brinda la posibilidad de establecer estrategias para contribuir al desarrollo y capacitación de profesionales. Objetivos. 1) Describir las características de las residencias de TIP del país. 2) Evaluar si existen características que se relacionen con una mayor ocupación de las vacantes. 3) Explorar la inserción laboral en el hospital formador de los residentes. Diseño. Descriptivo, observacional. Encuesta nacional. Criterios de inclusión. Residencias de TIP funcionales entre el 1/4/2014 y el 31/5/2014. Resultados. Se analizaron 31 residencias. Solo 11/31 tenían volumen de internación anual >400 pacientes. No había normas y/o criterios de atención en 9/31. En 17/31, el programa estuvo adecuado al marco de referencia nacional. Hubo 13/31 que no contaban con jefe ni instructor de residentes. Fueron acreditadas por el Ministerio de Salud 5/31. Hubo 65 vacantes; el número aumentó en los últimos 4 años; la ocupación disminuyó de 59% en 2009 a 30% en 2013. El 60% de los residentes tuvo inserción laboral en la TIP formadora. El análisis de regresión logística multivariado identificó la variable ingresos anuales > 400 pacientes como predictora independiente de ocupación de vacantes > 60%. Conclusiones. 1) Hay un déficit en la ocupación de cargos. 2) El número de residencias acreditadas es escaso. 3) Las unidades de cuidados intensivos pediátricos con mayor número de ingresos se asociaron a una mayor cobertura de vacantes. 4) Más de la mitad de los residentes se insertaron laboralmente en la TIP formadora.

Introduction. Pediatric intensive care residency programs have been in place in Argentina for just a few years. Knowing their status offers the possibility to establish strategies to help with professional development and training. Objectives. 1) To describe the characteristics of pediatric intensive care residency programs across Argentina. 2) To assess whether certain characteristics are related to a higher vacancy filling rate. 3) To assess job placement in the hospital where residents are trained. Design. Descriptive, observational study. National survey. Inclusion criteria. Pediatric intensive care residency programs in place between April 1st, 2014 and May 31st, 2014. Results. Thirty-one residency programs were analyzed. Only 11/31 had an annual hospitalization volume >400patients. There were no guidelines and/or criteria for care in 9/31. The program suited the national reference frameworkin17/31. There was no head ofresidents or resident trainer in 13/31. Only 5/31 had been certified by the Ministry of Health. There were 65 vacancies; this number increased in the past four years; vacancy filling rate decreased from 59% in 2009 to 30% in 2013. Sixty percent of residents got a job in the pediatric intensive care unit where they were trained. A multivariate logistic regression analysis identified the outcome measure annual hospitalization volume >400 patients as an independent predictor of vacancy filling rate >60%. Conclusions. 1) Vacancy filling is deficient. 2) The number of certified residency programs is scarce. 3) Pediatric intensive care units with a higher number of hospitalizations were associated with a higher vacancy filling rate. 4) More than half of residents got a job in the pediatric intensive care unit where they were trained.

Assessment of apple core collections constructed using phenotypic and genotypic data.

Several types of information can be used to select core collections, including passport data, agronomic data, and molecular data. However, little is known about the ability of core collections to retain the genetic diversity and structure of the whole collection for characters that were not considered during the selection, particularly when molecular markers are used. In this study, two core subsets were established for the apple (Malus spp) germplasm bank curated at the Apple Research Station, National Institute of Horticultural and Herbal Science, Korea, based upon genetic diversity estimated with 14 simple sequence repeat markers, and phenotypic diversity based on 23 traits. Comparisons between these two subsets and with the whole collection were used to determine the effect of the data used in the selection on phenotypic and genetic diversity, and population structure. The two subsets had a similar diversity and did not differ from the original collection, according to the Nei and Shannon diversity indices. Allele and class frequencies were also maintained in the two subsets. Overall, the type of data used to construct the core collection had little influence on the phenotypic and genetic diversity retained. Therefore, in the case of apple collections, the use of molecular markers is preferable, because they allow rapid and reliable characterization.

Ethylene-dependent regulation of an α-L-arabinofuranosidase is associated to firmness loss in 'Gala' apples under long term cold storage.

Fruit texture changes impair the quality of apples submitted to long term storage, especially under cold. The changes are due to cell wall modifications during ripening and senescence and are associated to ethylene. We have investigated the activity of α-l-arabinofuranosidase, a glycosyl hydrolase acting on the side chains of pectin in the cell wall and middle lamella. The transcription of arabinofuranosidase coding sequences 1 and 3 was investigated in plant organs and in response to ethylene, employing hormone application and 1-methylcyclopropene. The transcription of arabinofuranosidase genes is not restricted to fruits, although upregulated by ripening and ethylene. Transcripts of the genes were detected under cold storage up to 180 days. Similarly, arabinofuranosidase activity increased with rising levels of ethylene and under cold storage. Levels of arabinofuranosidase3 transcripts were higher than those of arabinofuranosidase1, suggesting that the first is an important contributor to enzyme activity and texture changes during cold storage.

Functional diversification of the dehydrin gene family in apple and its contribution to cold acclimation during dormancy.

Dehydrins (DHN) are proteins involved in plant adaptive responses to abiotic stresses, mainly dehydration. Several studies in perennial crops have linked bud dormancy progression, a process characterized by the inability to initiate growth from meristems under favorable conditions, with DHN gene expression. However, an in-depth characterization of DHNs during bud dormancy progression is still missing. An extensive in silico characterization of the apple DHN gene family was performed. Additionally, we used five different experiments that generated samples with different dormancy status, including genotypes with contrasting dormancy traits, to analyze how DHN genes are being regulated during bud dormancy progression in apple by real-time quantitative polymerase chain reaction (RT-qPCR). Duplication events took place in the diversification of apple DHN family. Additionally, MdDHN genes presented tissue- and bud dormant-specific expression patterns. Our results indicate that MdDHN genes are highly divergent in function, with overlapping levels, and that their expressions are fine-tuned by the environment during the dormancy process in apple.