ABSTRACT
As one of the key enzymes in cell metabolism, the activity of citrate synthase 3 (CS3) regulates the substance and energy metabolism of organisms. The protein members of CS3 family were identified from the whole genome of apple, and bioinformatics analysis was performed and expression patterns were analyzed to provide a theoretical basis for studying the potential function of CS3 gene in apple. BLASTp was used to identify members of the apple CS3 family based on the GDR database, and the basic information of CS3 protein sequence, subcellular localization, domain composition, phylogenetic relationship and chromosome localization were analyzed by Pfam, SMART, MEGA5.0, clustalx.exe, ExPASy Proteomics Server, MEGAX, SOPMA, MEME, WoLF PSORT and other software. The tissue expression and inducible expression characteristics of 6 CS3 genes in apple were determined by acid content and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Apple CS3 gene family contains 6 members, and these CS3 proteins contain 473-608 amino acid residues, with isoelectric point distribution between 7.21 and 8.82. Subcellular localization results showed that CS3 protein was located in mitochondria and chloroplasts, respectively. Phylogenetic analysis divided them into 3 categories, and the number of genes in each subfamily was 2. Chromosome localization analysis showed that CS3 gene was distributed on different chromosomes of apple. The secondary structure of protein is mainly α-helix, followed by random curling, and the proportion of β-angle is the smallest. The 6 members were all expressed in different apple tissues. The overall expression trend from high to low was the highest relative expression content of MdCS3.4, followed by MdCS3.6, and the relative expression level of other members was in the order of MdCS3.3 > MdCS3.2 > MdCS3.1 > MdCS3.5. qRT-PCR results showed that MdCS3.1 and MdCS3.3 genes had the highest relative expression in the pulp of 'Chengji No. 1' with low acid content, and MdCS3.2 and MdCS3.3 genes in the pulp of 'Asda' with higher acid content had the highest relative expression. Therefore, in this study, the relative expression of CS3 gene in apple cultivars with different acid content in different apple varieties was detected, and its role in apple fruit acid synthesis was analyzed. The experimental results showed that the relative expression of CS3 gene in different apple varieties was different, which provided a reference for the subsequent study of the quality formation mechanism of apple.
Subject(s)
Citric Acid , Malus/genetics , Citrate (si)-Synthase , Phylogeny , CitratesABSTRACT
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.
Subject(s)
Malus/genetics , Ascorbic Acid , Fruit/genetics , Genotype , AntioxidantsABSTRACT
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'.
Subject(s)
Malus/growth & development , Malus/genetics , Fruit , Crop ProductionABSTRACT
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.
Subject(s)
Malus/anatomy & histology , Malus/growth & development , Malus/genetics , Malus/chemistryABSTRACT
The environmental gas concentration affects the storage period and quality of fruits and vegetables. High concentration CO₂ treating for a long time will cause damage to fruits, However, the specific molecular mechanism is unclear. To analyze the mechanism of CO₂ injury in apple, high-throughput sequencing technology of Illumina Hiseq 4000 and non-targeted metabolism technology were used to analyze the transcriptome sequencing and metabolomics analysis of browning flesh tissue of damage fruit and normal pulp tissue of the control group. A total of 6 332 differentially expressed genes were obtained, including 4 187 up-regulated genes and 2 145 down regulated genes. Functional analysis of the differentially expressed genes confirmed that the occurrence of CO₂ injury in apple was related to redox process, lipid metabolism, hormone signal transduction process and energy metabolism process. Twenty candidate browning genes were successfully screened, among which grxcr1 (md14g1137800) and gpx (md06g1081300) participated in the reactive oxygen species scavenging process, and pld1_ 2 (md15g1125000) and plcd (md07g1221900) participated in phospholipid acid synthesis and affected membrane metabolism. mdh1 (md05g1238800) participated in TCA cycle and affected energy metabolism. A total of 77 differential metabolites were obtained by metabolomic analysis, mainly organic acids, lipids, sugars and polyketones, including 35 metabolites related to browning. The metabolism of flavonoids was involved in the browning process of apple. Compared with the control tissue, the content of flavonoids such as catechin and quercetin decreased significantly in the damaged apple tissue, the antioxidant capacity of cells decreased, the redox state was unbalanced, and the cell structure was destroyed, resulting in browning. The results of this study further enrich the theoretical basis of CO₂ damage, and provide reference for the practical application of high concentration CO₂ preservation technology.
Subject(s)
Carbon Dioxide , Fruit , Gene Expression Regulation, Plant , Malus/genetics , Metabolome , TranscriptomeABSTRACT
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.
Subject(s)
Malus/genetics , Malus/virology , Flexiviridae , Seed Bank , Enzyme-Linked Immunosorbent Assay , Reverse Transcriptase Polymerase Chain Reaction , Malus/growth & developmentABSTRACT
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.
Subject(s)
Cloning, Molecular , Dioxygenases/genetics , Fruit/genetics , Gene Expression , Malus/genetics , Plant Proteins/genetics , Stress, Physiological/genetics , Amino Acid Sequence , Chromosome Mapping , Dioxygenases/chemistry , Fruit/growth & development , Gene Expression Regulation, Plant , Introns , Molecular Sequence Data , Malus/classification , Malus/growth & development , Phylogeny , Promoter Regions, Genetic , Plant Proteins/chemistry , Regulatory Sequences, Nucleic Acid , Sequence Alignment , Sequence Analysis, DNAABSTRACT
Malus zumi is known as an excellent dwarfing apple rootstock occurring in natural or arid/semiarid soil or salina. Gene manipulation of M. zumi through transgenic technology can modify plant feature for further improvement fruit tree production by grafting the scion on a transgenic rootstock. Here, we report the establishment of an efficient, in vitro, shoot regeneration system and Agrobacterium tumefaciens- mediated transformation from the leaf explants for Malus zumi (Matsumura) Rehd. Leaf explants were infected with Agrobacterium strains containing nptII and gus gene. The highest frequency of shoot regeneration was obtained on MS medium containing 500 mg l-1 Lactalbumin hydrolysate, 30 g l-1 fructose, supplemented with 3.0 mg l-1 BA, 0.2 mg l-1 NAA.Using fructose instead of sucrose significantly increases the shoot regeneration and decreases vitrification. This regeneration procedure was incorporated into an Agrobacterium-mediated transformation procedure in M. zumi. Kanamycin was an efficient selective agent for selection. Pre-selection (5 days after co-cultivation) improved the transformation efficiency. The emergence of expected bands by PCR analysis and Southern blot in transgenic plantlets confirmed the transformation of foreign DNA into plant genome.
Subject(s)
Malus/genetics , Plants, Genetically Modified , Agrobacterium tumefaciens/enzymology , Agrobacterium tumefaciens/metabolism , Blotting, Southern , Kanamycin/pharmacokinetics , Kanamycin/chemical synthesis , Kanamycin/therapeutic use , Polymerase Chain ReactionABSTRACT
The generation of transgenic apple plants relies on the molecular analysis of transgene integration and expression based on polymerase chain reaction (PCR) analysis, blotting techniques and enzymatic assays on vitro leaves of putative transgenic regenerates. In order to assess the uniformity and the stability of transfer DNA (T-DNA) integration and gene expression, we studied 26 transgenic apple lines carrying the attacin E gene from Hyalophora cecropia, the beta-glucuronidase gene, and the nptII gene. Plants were evaluated using standard molecular techniques, such as PCR, Southern blot, reverse transcription PCR (RT-PCR) and Enzyme Linked Immunosorbent Assay (ELISA), and propagated in vitro on non-selective antibiotic-free media for four years to mimic natural conditions in the field. In some T-lines transgene integration and expression did not remain stable; differences were also found between distinct plants of a single T-line. Individual plants with partially or completely silenced transgenes were identified as well as plants with non-detectable T-DNA. Several lines appeared chimeric or partially silenced. Although most molecular techniques can reliably detect the presence of transgenic cells, they often fail to detect mixtures of transformed and non-transformed cells, or cells with silenced transgenes. This should be taken into consideration, especially in the case of vegetatively propagated trees, where non-transformed or silenced plant parts could mistakenly be used as propagation material.