Genome-wide analysis of G-type lectin genes in Fragaria vesca and functional characterization of FaMBL1 gene in defense response of F. × ananassa to fungal pathogens

Strawberry (Fragaria × ananassa) is an important soft fruit but easily to be infected by pathogens. Anthracnose and gray mold are two of the most destructive diseases of strawberry which lead to serious fruit rot. The first chapter introduced strawberry anthracnose caused by Colletotrichum acutatum. The infection strategy, disease cycle and management of C. acutatum on strawberry were reported. Likewise, the second chapter summarized the infection strategy of Botrytis cinerea and the defense responses of strawberry. As we already know white unripe strawberry fruits are more resistant to C. acutatum than red ripe fruits. During the interaction between strawberry white/red fruit and C. acutaum, a mannose binding lectin gene, FaMBL1, was found to be the most up-regulated gene and induced exclusively in white fruit. FaMBL1 belongs to the G-type lectin family which has important roles in plant development and defense process. To get insight into the role of FaMBL1, genome-wide identification was carried out on G-type lectin gene family in Fragaria vesca and the results were showed in chapter 3. G-type lectin genes make up a large family in F. vesca. Active expression upon biotic/abiotic stresses suggested a potential role of G-lectin genes in strawberry defenses. Hence, stable transgenic strawberry plants with FaMBL1 gene overexpressed were generated. Transformed strawberry plants were screened and identified. The results were showed in chapter 4, content of disease-related phytohormone, jasmonic acid, was found decreased in overexpressing lines compared with wild type (WT). Petioles inoculated by C. fioriniae of overexpressing lines had lower disease incidence than WT. Leaves of overexpressing lines challenged by B. cinerea showed remarkably smaller lesion diameters compared with WT. The chitinase 2-1 (FaChi2-1) showed higher expression in overexpressing lines than in WT during the interaction with B. cinerea, which could be related with the lower susceptibility of overexpressing lines.

Isolation of micropropagated strawberry endophytic bacteria and assessment of their potential for plant growth promotion

Twenty endophytic bacteria were isolated from the meristematic tissues of three varieties of strawberry cultivated in vitro, and further identified, by FAME profile, into the genera Bacillus and Sphingopyxis. The strains were also characterized according to indole acetic acid production, phosphate solubilization and potential for plant growth promotion. Results showed that 15 strains produced high levels of IAA and all 20 showed potential for solubilizing inorganic phosphate. Plant growth promotion evaluated under greenhouse conditions revealed the ability of the strains to enhance the root number, length and dry weight and also the leaf number, petiole length and dry weight of the aerial portion. Seven Bacillus spp. strains promoted root development and one strain of Sphingopyxis sp. promoted the development of plant shoots. The plant growth promotion showed to be correlated to IAA production and phosphate solubilization. The data also suggested that bacterial effects could potentially be harnessed to promote plant growth during seedling acclimatization in strawberry.

Chemical Composition and Antioxidant/Antidiabetic Potential of Brazilian Native Fruits and Commercial Frozen Pulps

Foods provide essential and bioactive compounds with health-promoting properties such as antioxidant, anti-inflammatory, and hypocholesterolemic activities, which have been related to vitamins A, C, and E and phenolic compounds such as flavonoids. Therefore, the aim of this work was to identify potential sources of bioactive compounds through the determination of flavonoids and ellagic acid contents and the in vitro antioxidant capacity and alpha-glucosidase and alpha-amylase inhibitory activities of Brazilian native fruits and commercial frozen pulps. Camu-camu, cambuci, uxi, and tucuma and commercial frozen pulps of cambuci, cagaita, coquinho azedo, and araca presented the highest antioxidant capacities. Cambuci and cagaita exhibited the highest alpha-glucosidase and alpha-amylase inhibitory activities. Quercetin and kaempferol derivatives were the main flavonoids present in most of the samples. Ellagic acid was detected only in umbu, camu-camu, cagaita, araca, and cambuci. According to the results, native Brazilian fruits can be considered as excellent sources of bioactive compounds.

Anthocyanin synthesis, growth and nutrient uptake in suspension cultures of strawberry cells

Strawberry (Fragaria ananassa cv. Shikinari) cell suspension cultures carried out in shake flasks for 18 d were closely examined for cell growth, anthocyanin synthesis and the development of pigmented cells in relation to the uptake of carbohydrate, extracellular PO4, NO3, NH4, and calcium. Cell viability, extracellular anthocyanin content, pH and electrical conductivity of the broth were also monitored. The specific growth rate of strawberry cells at exponential phase was 0.27 and 0.28 d(-1) based on fresh and dry weight, respectively. Anthocyanin synthesis was observed to increase continuously to a maximum value of 0.86 mg/g fresh cell weight (FCW) at day 6, and was partially growth-associated. Anthocyanin synthesis was linearly related to the increase in pigmented cell ratio, which increased with time and reached a maximum value of ca. 70% at day 6 due to reduction in cell viability and depletion of substrate. Total carbohydrate uptake was closely associated with increase in cell growth, and glucose was utilized in preference to fructose. Nitrate and ammonia were consumed until 9 d of culture, but phosphate was completely absorbed within 4 d. Calcium was assimilated throughout the growth cycle. After 9 d, cell lysis was observed which resulted in the leakage of intracellular substances and a concomitant pH rise. Anthocyanin was never detected in the broth although the broth became darkly pigmented during the lysis period. This suggests that anthocyanin was synthesized only by viable pigmented cells, and degraded rapidly upon cell death and lysis. Based on the results of kinetic analysis, a model was developed by incorporating governing equations for the ratio of pigmented cells into a Bailey and Nicholson's model. This was verified by comparison with the experimental data. The results suggest Bat the model satisfactorily describes the strawberry cell culture process, and may thus be used for process optimization.

Produção e qualidade de propágulos de morangueiro em diferentes concentrações de nitrogênio no cultivo sem solo

O objetivo deste trabalho foi avaliar o efeito da concentração de nitrogênio na produção e na qualidade de mudas de raízes nuas e pontas de estolões de morangueiro no cultivo sem solo. O plantio das matrizes foi realizado em 13 de setembro de 2007, e a colheita das mudas em 18 de fevereiro de 2008. Os tratamentos foram quatro concentrações de N na solução nutritiva de 8, 11, 14 e 17 mmol L-1. O delineamento utilizado foi o inteiramente casualizado, com cinco repetições de 3,6 m². No momento da colheita foram determinados o número e o diâmetro da coroa das mudas de raízes nuas e o número de pontas de estolões. As diferentes concentrações de N não afetaram o número de mudas e de pontas de estolões, cujas médias foram de 339 e 836, respectivamente. O diâmetro da coroa das mudas de raízes nuas diminuiu linearmente com o aumento da concentração de N na solução nutritiva. Concluiu-se que o aumento da concentração de N na solução nutritiva, no sistema de cultivo sem solo com substrato de areia, não afeta o número de mudas de raízes nuas e nem de pontas de estolões, mas reduz o diâmetro da coroa das mudas de raízes nuas de morangueiro.

Divergência genética em cultivares de morangueiro, baseada em caracteres morfoagronômicos

Os caracteres morfoagronômicos são tradicionalmente usados na caracterização de cultivares e no estudo da divergência genética, contribuindo na definição de estratégias para o melhoramento genético. Este trabalho teve por objetivo avaliar a divergência genética por meio de caracteres morfoagronômicos de 11 cultivares de morangueiro (Aromas, Camarosa, Camino Real, Campinas, Diamante, Dover, Oso Grande, Sweet Charlie, Toyonoka, Tudla e Ventana), nas condições climáticas da região Centro-Sul do Paraná. Foram analisados 29 caracteres morfoagronômicos relacionados com a planta, folha, flor, fruto e aquênios do morangueiro. As similaridades genéticas foram calculadas por meio de análise multivariada e, os cultivares, agrupados com base na matriz de similaridade genética, usando-se UPGMA. Dentre os 29 caracteres morfoagronômicos avaliados, oito apresentaram diferenças não significativas (p < 0,05). A similaridade média foi de 38%, variando de 19 (aromas e camino real) a 62% (Camino Real e Camarosa; Aromas e Sweet Charlie). O dendrograma alocou os cultivares em quatro grupos, contudo, essa divisão não foi coerente com a origem e genealogia dos cultivares. O cultivar Tudla apresenta elevado potencial "per se" para utilização em programas de melhoramento. O cruzamento mais promissor com base nos caracteres morfoagronômicos é entre os cultivares Camarosa e Campinas.

Post-harvest conservation of organic strawberries coated with cassava starch and chitosan

The strawberry is as non-climacteric fruit, but has a high post-harvest respiration rate, which leads to a rapid deterioration at room temperature. This study aimed to evaluate the application of biodegradable coating on postharvest conservation of organic strawberries, cv. Camarosa, packed in plastic hinged boxes and stored at 10ºC. The treatments consisted of: a) control; b) 2% cassava starch; c) 1% chitosan; and d) 2% cassava starch + 1% chitosan. Physical and chemical characteristics of fruits were evaluated at 3, 6 and 9 days of storage, and microbiological and sensory analyses were carried out at the end of the storage period. The treatments influenced positively the post-harvest quality of organic strawberries. The coating cassava starch + chitosan provided the best results, with less than 6% of loss in fruit mass, lower counts of yeast and psychrophilic microorganisms and the best appearance according to the sensory analysis.

Produção de cultivares de morango, utilizando túnel baixo em Pelotas

O objetivo deste trabalho foi avaliar o desempenho produtivo de novos cultivares de dia-curto de morangueiro ('Earlibrite', 'Sabrosa' e 'Florida Festival'), nas condições edafo-climáticas de Pelotas (RS), tendo como padrão o cultivar 'Camarosa', usando-se túnel baixo. O experimento foi realizado com sistema de irrigação por gotejamento. O transplantio das mudas foi realizado em maio de 2008, sendo utilizadas mudas procedentes da região da Patagônia, na Argentina. O delineamento experimental foi de blocos casualizados, com parcelas subdivididas no tempo, com quatro repetições. As unidades experimentais foram constituídas por 42 plantas, que foram avaliadas, semanalmente, quanto ao número, massa da matéria fresca e produção de frutos, no período de agosto a dezembro de 2008. Nas avaliações, consideraram-se apenas os frutos comercializáveis, descartando-se aqueles com defeitos e com massa inferior a 5 g. Verificou-se que a produção acumulada de frutos por planta do 'Sabrosa' é maior que a do 'Florida Festival' e do 'Earlibrite', menor, porém, que a do 'Camarosa'. O número de frutos produzidos por planta do 'Sabrosa' e do 'Florida Festival' é maior que o do 'Earlibrite', menor, porém, que o do 'Camarosa'. Quanto à produção de massa da matéria fresca média dos frutos, o 'Sabrosa' e o 'Camarosa' são superiores a do 'Florida Festival' e a do'Earlibrite'. Apesar de apresentarem padrões de desempenho produtivo diferenciados, os três novos cultivares de morangueiro testados mostraram-se com alto potencial produtivo, constituindo-se em interessantes alternativas de produção para os agricultores de Pelotas.

Biochemical and morpho-anatomical analyses of strawberry vitroplants hyperhydric tissues affected by BA and gelling agents

In vitro propagation has become an effective practice for large-scale production of strawberry plants. The objective of this study was to evaluate the hyperhydricity and the multiplication capacity of two strawberry varieties (Fragaria x ananassa Duch. 'Dover' and 'Burkley') propagated in vitro. Plants maintained in MS medium supplemented with 1.0 mg L-1 BA were individualized and transferred to the same medium solidified with Agar (6.5 g L-1) or Phytagel® (2.5 g L-1) and BA at different concentrations (0; 0.5; 1.0; 2.0 and 3.0 mg L-1). Biochemical and anatomical analyses were carried out, as well as the analysis of the morphological hyperhydricity characteristics. The analysis of data showed: a) the increase in cytokinin concentration increased hyperhydricity frequency in both varieties; b) at concentrations up to 2.0 mg L-1 BA, the replacement of Agar by Phytagel® induced a higher formation of hyperhydric shoots; and c) the addition of BA induced oxidative stress, which is characterized by increased antioxidant activity and lipid peroxidation, as well as alterations at the cellular level, such as malformation of stomata and epidermal cells. In conclusion, the culture medium containing 0.5 mg L-1 BA solidified with Agar provided lower hyperhydricity percentages in association with higher rates of shoot proliferation in strawberry.

Heat shock and salicylic acid on postharvest preservation of organic strawberries

Heat shock and salicylic acid have been studied on shelf-life extension of fruits. The benefits of these techniques have been related to their effect on inducing physiological defense responses against the oxidative stress and pathogen development. The objective of this study was to evaluate the effect of heat shock and salicylic acid on the postharvest preservation and contents of total phenolics, anthocyanins, ascorbic acid, fresh weight loss and microbiological quality of organic strawberries cv. Dover. Strawberries produced organically and stored at 5 ºC were subjected to heat shock (45 ºC ± 3 ºC for 3 h), application of salicylic acid (soaking in 2.0 mmol L-1 solution), heat shock in combination with salicylic acid and control. After treatment, the fruits were packed and stored in a climatic chamber at 5 ºC ± 2 ºC. At 1, 7 and 14 days, the experimental units were removed from refrigeration and kept at room temperature of approximately 20 ºC for two days. There was no effect of treatments on fresh weight loss, incidence of pathogens or chemical variations in strawberry fruits during the storage period. In natural conditions, organically grown strawberries remained in good condition for sale up to seven days of storage in all treatments.

Efeitos de reguladores de crescimento nas características organográficas e na produtividade do morangueiro 'Sequóia'

Este estudo teve por objetivo verificar a possibilidade de se melhorar a produção de pseudofrutos de morangueiro 'Sequóia' e de promover alterações morfológicas nas plantas tratadas com fitoreguladores. O experimento instalado no campo e delineado em blocos casualizados, constou de 5 tratamentos e 4 blocos, com 12 plantas por parcela. Além do controle, foram testados os reguladores vegetais ácido giberélico (GA3) e ácido naftalenoacético NAA e os bioestimulantes Ergostim e Atonik. A dose total foi de 30ppm, parcelada em 3 aplicações, a intervalos semanais, iniciadas após o inicio do florescimento. Não houve diferenças significativas entre o controle e os demais tratamentos quanto aos parâmetros organográficos estudados, embora a conformação das plantas tratadas com GA3 tenha sido alterada. A análise dos dados de produção não revelaram diferenças significativas entre o controle e os tratamentos, apesar do número de morangos produzidos pelo GA3 e pelo controle tenha sido superior ao dos demais. O peso, o comprimento e o diâmetro dos pseudofrutos não foram alterados pelos tratamentos. Foi utilizado o modelo estatístico "split-plot", a fim de detectar a interação entre os tratamentos e as épocas de produção. As épocas de produção interferiram no número e tamanho de morangos nos diferentes tratamentos. Atonik induziu mo rangos com teores mais elevados de açúcares solúveis em relação ao controle e ao Ergostim.

Biologia e tabela de vida de fertilidade de Frankliniella occidentalis (Pergande) (Thysanoptera, Thripidae) em morangueiro

O objetivo deste trabalho foi estudar aspectos biológicos de Frankliniella occidentalis (Pergande) (Thysanoptera, Thripidae), considerando que no Brasil quase nada se sabe sobre a fauna de tripes associada à cultura do morangueiro. Larvas recém-eclodidas foram individualizadas em placas de Petri contendo uma flor ou um folíolo de morangueiro, mantidas em câmaras climatizadas (25 ± 1 ºC; 60 ± 10% U.R.; fotofase de 12 horas) e observadas diariamente até a morte. A duração média do período de larva-adulto e a viabilidade não diferiram entre os insetos mantidos em flores (8,49 ± 0,18 e 68,52%) e folíolos (8,85 ± 0,15 e 75,47%). A fecundidade média diária e a total foram mais elevadas quando flores foram fornecidas como alimento (7,4 ± 0,69 e 70,0 ± 9,18 ovos/fêmea respectivamente), em comparação com folíolos (2,4 ± 0,35 e 8,5 ± 1,13 ovos/fêmea, respectivamente). A duração média, em dias, do período embrionário foi distinta entre os indivíduos mantidos em flores (3,7 ± 0,03) e em folíolos (4,4 ± 0,09). A viabilidade dos ovos depositados sobre flores e folíolos foi de 65,5 ± 0,01 e 74,3 ± 0,03%, respectivamente. Com base na tabela de vida de fertilidade, o desempenho dos indivíduos de F. occidentalis que se desenvolveram em flores foi melhor, com uma geração (ovo-adulto) completada a cada 20,92 dias, a 25 °C.

Species of thrips (Insecta, Thysanoptera) in two strawberry production systems in Rio Grande do Sul State, Brazil

Species of thrips (Insecta, Thysanoptera) in two strawberry production systems in Rio Grande do Sul, Brazil. Thrips are tiny insects responsible for the reduction of strawberry fruit quality. The work aimed to record and quantify the thysanopterofauna present in two strawberry production systems, low tunnel and semi-hydroponic. Leaves, flowers and fruits were collected weekly, from July 2005 to December 2006 in Caxias do Sul and Bom Princípio municipalities, RS. A total of 664 individuals were collected, representing two families, four genus and 10 species: Frankliniella occidentalis (Pergande, 1895), F. schultzei (Trybom, 1910), F. rodeos Moulton, 1933, F. simplex (Priesner, 1924), F. williamsi (Hood, 1915), F. gemina (Bagnall, 1919), Frankliniella sp., Thrips tabaci (Lindeman, 1888), Thrips tabaci (Lindeman, 1888), Caliothrips fasciatus (Pergande 1895) from Thripidae and Heterothrips sp. from Heterothripidae. Frankliniella occidentalis represented 89.7% of the samples with 95.8% of the species collected in flowers, 3.9% in fruits and 0.8% in leaves. The results show that flowers are the most important food resource for these insects on strawberry plants. Frankliniella rodeos, F. simplex, F. williamsi, C. fasciatus, and Heterothrips sp. are new records on strawberry for Brazil.

The effects of a non-woven cover in combination with different soil mulches in strawberry cultivation

Selostus: Katteiden käyttö mansikanviljelyssä

Characterization of soil chemical properties of strawberry fields using principal component analysis

One of the largest strawberry-producing municipalities of Rio Grande do Sul (RS) is Turuçu, in the South of the State. The strawberry production system adopted by farmers is similar to that used in other regions in Brazil and in the world. The main difference is related to the soil management, which can change the soil chemical properties during the strawberry cycle. This study had the objective of assessing the spatial and temporal distribution of soil fertility parameters using principal component analysis (PCA). Soil sampling was based on topography, dividing the field in three thirds: upper, middle and lower. From each of these thirds, five soil samples were randomly collected in the 0-0.20 m layer, to form a composite sample for each third. Four samples were taken during the strawberry cycle and the following properties were determined: soil organic matter (OM), soil total nitrogen (N), available phosphorus (P) and potassium (K), exchangeable calcium (Ca) and magnesium (Mg), soil pH (pH), cation exchange capacity (CEC) at pH 7.0, soil base (V%) and soil aluminum saturation(m%). No spatial variation was observed for any of the studied soil fertility parameters in the strawberry fields and temporal variation was only detected for available K. Phosphorus and K contents were always high or very high from the beginning of the strawberry cycle, while pH values ranged from very low to very high. Principal component analysis allowed the clustering of all strawberry fields based on variables related to soil acidity and organic matter content.