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  • 1
    ISSN: 1432-2048
    Keywords: Key words: Anthracnose ; Disease resistance (grapevine) ; Proembryogenic mass ; Vitis (disease resistance)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract.  Proembryogenic masses of grapevine (Vitis vinifera L.) `Chardonnay' (clone 02Ch) were exposed to the culture filtrate of Elsinoe ampelina (deBary) Shear, the causal agent of anthracnose disease. After four or five cycles of recurrent in-vitro selection with medium containing 40% fungal culture filtrate, putative resistant lines RC 1 and RC 2 respectively, were established. The selected lines inhibited the growth of E. ampelina and Fusarium oxysporium (Schlecht.) (isolated from watermelon) in a dual-culture assay and reduced the growth of mycelium on a conditioned-medium test, thus suggesting the involvement of extracellular compounds in resistance. Sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) gel electrophoresis of extracellular proteins from spent suspension-culture medium showed enhanced secretion of new proteins by selected lines. A 36-kDa protein was immunodetected by a chitinase antiserum. This chitinase continued to express constitutively in differentiated somatic embryos and also in the intercellular fluids of plants regenerated from the selected lines. Somatic embryos from selected lines grew uninhibitedly in a medium containing 40% fungal culture filtrate, whereas non-selected (control) somatic embryos became necrotic and died within a few days. Plants regenerated from selected lines exhibited resistance to infection by E. ampelina in both greenhouse tests and detached leaf bioassays. Results suggest that embryogenic cells can be selected for resistance following in-vitro selection, resulting in resistant plants. Whether or not resistant cells pre-existed in the original embryogenic culture or were induced by the selection pressure could not be determined.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5044
    Keywords: Arachis species ; nurse culture ; plant regeneration ; protoplasts ; tissue culture ; wild peanut
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient protocol has been developed for protoplast culture and plant regeneration from wild peanut (A. paraguariensis) using a nurse culture method. Protoplasts were isolated from suspension cultures initiated from leaf-derived callus, imbedded in agarose blocks and co-cultured with nurse cells of the same species. Up to 10% of the protoplasts divided and formed compact callus colonies. The protoplast plating efficiency was correlated with both the length of the nurse cell co-cultivation period and the protoplast plating density. The optimal nurse culture duration was 14 d. The optimal plating density was 2×104 protoplasts/ml plating medium. Multiple shoots (up to 10 shoots per colony) were readily regenerated from protoplast-derived callus after transfer of callus to semi-solid modified MS medium containing 0.5 mg l-1 NAA and 1 mg l-1 BA. Plantlets with normal leaflets were obtained by rooting shoots on porous rootcubes saturated with modified MS medium containing 1 mg l-1 NAA.
    Type of Medium: Electronic Resource
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