Enzymatic isolation of living embryo sacs of Petunia
| Autor: | J. L. Van Went, H. S. Kwee |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1990 |
| Předmět: |
chemistry.chemical_classification
Laboratorium voor Plantencelbiologie Chromatography biology Embryo sac Enzymatic maceration Embryo Cell Biology Plant Science Petunia hybrida Protoplast biology.organism_classification Petunia Laboratory of Plant Cell Biology Enzyme chemistry embryonic structures Botany medicine Maceration (wine) Isolation technique Liberation Mannitol Ovule medicine.drug |
| Zdroj: | Sexual Plant Reproduction 3 (1990) Sexual Plant Reproduction, 3, 257-262 |
| ISSN: | 0934-0882 |
| DOI: | 10.1007/bf00202883 |
| Popis: | A 20%–25% yield of isolated and living embryo sacs of Petunia hybrida L. was obtained using an enzymatic maceration mixture containing 3% driselase (soluble fraction only), 0.1% MES buffer, pH 5.5, and 8% mannitol. For each maceration ± 450 ovules were incubated in 1 ml enzyme solution for 2 h at 30° C in a shaking waterbath (150 rpm). Subsequently, the enzyme solution was replaced by Brewbaker and Kwack's medium, pH 6.5, supplemented with 10% mannitol (BKM). Gentle agitation of the suspension resulted in the liberation of embryo sacs, which were then collected with a micropipette using a dissecting microscope and transferred to fresh BKM. The embryo sacs isolated are intact and living, and have maintained their original shape and organization When stored in BKM at room temperature the isolated embryo sacs remain alive for 8 h. Storage at 4° C results in a prolongation of viability of up to 80 h. Prolonged incubation of ovules or reincubation of isolated embryo sacs in the maceration mixture results in the liberation of the gametophytic cells as individual, living protoplasts. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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