Resumo -Os objetivos deste trabalho foram caracterizar a resistência à vassoura-de-bruxa de plantas de cacau originadas do cruzamento entre TSH 1188 e CCN 51 (população segregante), por meio de dois métodos de inoculação em condições de campo, e identificar marcadores microssatélites específicos para grupos de plantas resistentes e suscetíveis. As plantas-controle avaliadas pelos métodos de inoculação natural e inoculação artificial em campo produziram os mesmos padrões de sintomas. As plantas da população segregante também coincidiram os padrões de sintomas em 90%, por esses dois métodos. O método de inoculação artificial em campo permite detectar falso-resistentes. Dos 18 pares de primers microssatélites amplificados, 15 foram polimórficos entre os genitores, e seis entre os grupos de plantas segregantes contrastantes quanto à resistência à vassoura-de-bruxa. Foram confirmadas três marcas previamente associadas a QTL (locos para características quantitativas) relacionados com a resistência à vassoura-de-bruxa, comuns a outras populações. Também foram identificados três novos QTL para esta característica, típicos desta população, o que comprova sua utilidade para o melhoramento genético do cacaueiro.Termos para indexação: Theobroma cacao, Moniliophthora perniciosa, inoculação, marcador molecular, QTL, melhoramento genético.
Microsatellite markers related to resistance of cocoa tree against witches'-broomAbstract -The objectives of this work were to evaluate cocoa tree resistance against witches'-broom, in plants originated from the crossing between TSH 1188 and CCN 51 (segregating population), by means of two methods of inoculation in field conditions, and to identify microsatellite markers specific for resistant and susceptible plants. The control plants bore identical symptoms as the plants of the segregating population in 90% of the cases under the two methods. The method of artificial inoculation in the field allows the detection of false resistance to the disease. Of the 18 pairs of microsatellite primers amplified, 15 were polymorphic between genitors and six were polymorphic between the two groups of plants evaluated for resistance to witches'-broom. Three previously characterized markers were confirmed as associated to QTL (quantitative trait loci) related to resistance to the witches'-broom, which is common in other populations. Three new QTL for this characteristic, typical of this population, were also identified, which proves the utility of this population for cocoa breeding.
Advances in DNA technology, such as marker assisted selection, detection of quantitative trait loci and genomic selection also require the isolation of DNA from a large number of samples and the preservation of tissue samples for future use in cacao genome studies. The present study proposes a method for the preservation of sample tissues for DNA extraction and for manual extraction of large number of samples using spheres. The integrity and concentration of the DNA by these methods were assessed and compared with conventional method using mortar. The best parameters in order to obtain a fine powder using spheres was the use of 4 lyophilized leaf disks (50 mg), a single steel ball of 6 mm in diameter, followed by 30 s of manual maceration. The quantity of DNA obtained was four times higher than the conventional method. The purity of the DNA obtained was satisfactory and proved to be amplifiable by PCR using SSR primers. The present approach is a reliable, rapid, simple and consistent DNA isolation method for cacao, compared to the conventional methods. The protocol greatly increases the efficiency of extraction and suggests an inexpensive and practical way of DNA isolation of cacao for large scale.
This paper presents the description and analysis of a didactic experience involving the participation of a university and a community school, developed as part of the National Science and Technology Week, at a public university in northeastern Brazil. For this purpose, the use of learning station rotation enabled innovation in the teaching of physiology integrated with biochemistry and health education contents. The didactic approach consisted of creating a learning circuit comprising seven stations. The central theme of the stations emphasized physiology, with specific foci on biochemistry and cardiorespiratory and endocrine health. Each station provided unique activities related to the central theme, including a station concerning digital technology in physiology. The school students were divided into small groups (6 or 7 people) that rotated through the stations, with a total of 81 students visiting each station. A qualitative assessment was performed using a Likert-scale questionnaire to measure the level of satisfaction of the students. It was found that this didactic approach increased the receptivity of the students to the contents, facilitated student-teacher dialogue, and provided an excellent tool for establishing an interface between the university and the community school. Overall, 76.5% of the students rated the activity as excellent.
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