ISHS Acta Horticulturae 652: I International Symposium on Grapevine Growing, Commerce and Research
ESTABLISHMENT OF EMBRYOGENIC CULTURES FROM GYNOECIA AND ANTHERS IN VITIS VINIFERA L. 'TOURIGA NACIONAL' |
| Authors: | A.L. Pinto-Sintra, N. Magalhães |
| Keywords: | Somatic embryogenesis, plant regeneration, embryogenic calli long-term culture, secondary somatic embryogenesis, grapevine. |
| Abstract:
Portugal's vineyards are dominated by native grape varieties, Touriga Nacional being an important V. vinifera red cultivar used in Port wine, table wine and good varietal wine production. Genetic engineering is an attractive option to grape improvement because individual genes can be transferred into elite genotypes like Touriga Nacional, to overcome problems like sensibility to abiotic or biotic stress. In order to obtain a long-term source of somatic embryos to be used in transgenesis, embryogenic cultures were induced in four callogenesis induction media (CIM1: 1 μM BAP + 9 μM 2,4-D + 17 μM IASP; CIM2: 1 μM BAP + 9 μM 2,4-D; CIM3: 9 μM BAP + 4,5 μM 2,4-D; CIM4: 9 μM BAP + 4,5 μM 2,4-D + 17 μM IASP) from gynoecia and anthers of three Touriga Nacional selected clones. After the transfer to regeneration medium (RM, with 13,3 μM BAP and 0,3 % activated charcoal), the cultures showed high variation in the presented answers, going from clusters of somatic embryos at different development states, isolated somatic embryos arising directly from necrosed explants or embryogenic calli covered on surface with globular embryos, until browning and necrosed material (few gynoecia, a lot of anthers and some calli). Using appropriated medium (MM with 2,3 μM 2,4-D), maintenance of embryogenic calli has been obtained. Secondary somatic embryogenesis was observed when isolated somatic embryos were cultured in MM. Seven lines of embryogenic cultures were established (white or slightly yellow and friable embryogenic callus covered on surface with globular embryos), from the three clones. Upon transferring the embryogenic calli to RM, new somatic embryos were produced. Whenever somatic embryos clusters were detected, they were isolated and cultured on a germination media (GM, with 4,4 M BAP). After germination, they were cultured separately in culture tubes with MS depleted of growth regulators. Although some morphological variation occurred, plant regeneration from these embryogenic lines was successful. | |
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