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Previous issue date: 2010-04-26 === CAPES === The culture of strawberry (Fragaria x ananassa Duch.) is an agricultural activity of great socio-economic importance. In recent years, its production has increased in Brazil due to the introduction of new cultivars and use of quality plants. The vegetative propagation by stolons, usual in the species of the genus Fragaria also provide a limited number of plants and can spread viral diseases. The production of healthy plants can be obtained by meristem culture. Been strawberry cultivars in vitro responsive, with this technique it is possible to obtain a greatest number of plants. However the increase in the number of subcultures may induce somaclonal variation. In order to expand the availability of healthy plants of strawberry with genetic identity guarantee, this study aimed to analyze, with the use of morphological and molecular markers, with precision, the number of subcultures that allows large scale multiplication of strawberry, without loss of genetic identity. Explants of the cultivars Aromas, Camarosa and Camino Real, at the Laboratory of Plant Tissue Culture, in a first step, was submitted to twelve cycles of subculture, and in the following year, explants of the same cultivars, from the same mother plants, were subcultured for three cycles. The acclimatization and evaluation of the plants in the field were conducted in a greenhouse at Horticulture Sector of the Instituto de Agronomia of UFRRJ. The experimental design was randomized blocks in a factorial scheme 3x2, with the three genotypes being a factor and the two subculture levels other factor. In the Laboratory of Molecular Genetics of the Instituto de Biologia of UFRRJ, extraction of DNA was proceeded from leaves of regenerated in vitro plantlets of the three cultivars submitted to three, five and twelve subcultures and of the mother plants that were not produced by tissue culture. During the in vitro multiplication, in two levels of subcultures, we observed a greater tendency of hyperhydricity in Camino Real cultivar and a higher frequency of callus production in ?Camarosa?. After the acclimatization, plants exposed to twelve subcultures showed a higher average height. However, the average root length was not significantly different between the two levels of subcultures. In the field evaluation, for other quantitative traits, there were no significant differences between the two levels of subcultures. Cultivars Camarosa and Camino Real showed the highest number of characteristics with a variation between the two levels of subcultures, with the greatest variations observed in plants subjected to three subcultures. Analysing the field qualitative characteristics, the variation observed between the third and 12th subcultures, was present only the frequency distribution of leaf brightness, length and width of terminal leaflet. === A cultura do morangueiro (Fragaria x ananassa Duch.) ? uma atividade agr?cola de grande import?ncia s?cio-econ?mica. Nos ?ltimos anos, o seu cultivo tem aumentado no Brasil devido ? introdu??o de novas cultivares, sendo a produ??o de mudas de qualidade um dos fatores que afetam a expans?o da produ??o. A propaga??o vegetativa de plantas do g?nero Fragaria al?m de fornecer um n?mero limitado de prop?gulos, pode disseminar doen?as vir?ticas e radiculares. A produ??o de mudas sadias pode ser obtida atrav?s da cultura de meristemas. Considerando que cultivares de morangueiro s?o responsivas a propaga??o in vitro, atrav?s desta t?cnica ? poss?vel obter o maior n?mero de plantas, aumentando o n?mero de subcultivos, contudo, este acr?scimo pode induzir a ocorr?ncia de varia??o somaclonal. A fim de ampliar a disponibilidade de mudas sadias de morangueiro e com garantia de identidade gen?tica, este trabalho teve como objetivo analisar, a partir do uso de marcadores morfol?gicos, o n?mero de subcultivos que permita a multiplica??o do morangueiro em larga escala, sem que ocorra a perda da identidade gen?tica dos clones submetidos a este processo. No Laborat?rio de Cultura de Tecidos Vegetais, explantes das cultivares Aromas, Camarosa e Camino Real, foram submetidos em uma primeira etapa a doze ciclos de subcultivos e, no ano seguinte, os explantes das mesmas cultivares provenientes das mesmas plantas matrizes foram subcultivados por tr?s ciclos. A aclimatiza??o e avalia??o das plantas a campo foram realizadas em estufa no Setor de Horticultura do Instituto de Agronomia da UFRRJ. O delineamento experimental foi em blocos casualizados em esquema fatorial 3x2, sendo um fator os tr?s gen?tipos e o outro, os dois n?veis de subcultivos. Durante a fase de multiplica??o in vitro, nos dois n?veis de subcultivos, foi observado um maior n?mero de frascos com brota??es com sintoma de hiperhidricidade na cultivar Camino Real e forma??o de calos na cultivar Camarosa. Ap?s a fase de aclimatiza??o foi observado que plantas submetidas a doze subcultivos apresentaram maior altura m?dia da parte a?rea, entretanto, para o comprimento m?dio da raiz n?o foi observada diferen?a significativa entre os dois n?veis de subcultivos. Na avalia??o a campo das demais caracter?sticas quantitativas, n?o foram observadas diferen?as significativas entre os dois n?veis de subcultivos. Com doze subcultivos in vitro de plantas de morangueiro, das cultivares Aromas, Camarosa e Camino Real, ? poss?vel obter maior n?mero mudas micropropagadas sem perda da estabilidade gen?tica. A cultivar
Camarosa apresentou valores m?dios superiores para altura da parte a?rea ap?s a fase de aclimatiza??o e a campo e, massas fresca e seca da parte a?rea, quando submetida a doze ciclos de subcultivos. As cultivares Camarosa e Camino Real apresentaram maior n?mero de caracter?sticas quantitativas com varia??o entre os dois n?veis de subcultivos, sendo as maiores varia??es observadas em plantas submetidas a tr?s subcultivos. A cultivar Camino Real seguida da ?Camarosa? apresentaram um maior n?mero de caracter?sticas qualitativas com varia??o na distribui??o das frequ?ncias entre os dois n?veis de subcultivos. A cultivar Aromas apresentou uma maior estabilidade gen?tica em rela??o ?s caracter?sticas quantitativas e qualitativas.
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