Reduction of the number of nuclei per cell ofHelminthosporium euphorbiae by protoplast isolation |
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Authors: | Tânia Aparecida da Silva Luzia Doretto Paccola-Meirelles |
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Institution: | (1) Departmento de Biologia Geral-Centro de Ciências Biológicas-Universidade Estadual de Londrina, Campus Universitário, Caixa Postal 6001, CEP 86051-970 Londrina, Pr. Brasil |
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Abstract: | Helminthosporium euphorbiae is a pathogen of the weedEuphorbia heterophylla, which causes severe losses in soybean (Glycine max) crops. The fungus causes leaf loss and affects germination, making it a promising biocontrol agent for this weed. In order
to start a breeding program for this species, four isolates were examined for number of nuclei in the conidia and hyphae and
nuclear behavior at different cultivation stages. The conidia were multinucleated with about 20 nuclei per conidium, and 5
to 7 nuclei were observed in the hyphae compartments. The high number of nuclei makes the genetic manipulation of this species
diffucult, so the protoplast formation is an alternative for obtaining cells with a reduced number of nuclei. Thus the experimental
conditions for the production and regeneration of protoplasts inH. euphorbiae were determined by assessing three enzymatic complexes and seven osmotic stabilizers. The efficiency of formation and regeneration
frequency of the protoplasts varied depending on isolates, stabilizers and enzyme mixture used. The number of nuclei estimated
per protoplast was reduced to 1 to 6, depending on the stage of mycelial growth during the protoplast formation process. |
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Keywords: | cytology genetics Helminthosporium euphorbiae protoplasts |
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