Method for extracting pear pollen tube vacuoles
A pollen tube and vacuole technology, applied in the field of plant cell biology, can solve the problems of large amount of materials, unsuitable application of pear pollen, difficulty in obtaining materials, etc., and achieve the effect of less pollution
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[0046] 1) Pollen collection: collect anthers at the big bud stage and dry them with silica gel.
[0047] 2) Pollen culture: separate the pollen from the dried anther, take 3 mg of pollen, add it to 2 ml of pollen medium, and add 0.005% neutral red dye. Under dark conditions, culture at 23-27°C, 140-180rpm, for 3-4 hours. The formula of pollen medium: 100g / L sucrose, 0.3g / L Ca(NO 3 ) 2 4H 2 O, 30mM MES (α-sodium sulfomethyl ester sulfonate), 0.1g / L boric acid were dissolved in distilled water and fixed to volume, and the pH value was adjusted to 5.9-6.4 with Tris (trishydroxymethylaminomethane).
[0048] 3) Release of protoplasts: mix the cultured pollen and extracellular fluid at a volume ratio of 1:3, add 1% cellulase, 1% isolated enzyme, 23-27°C, under dark conditions, 60- 80rpm, react for 10-15 minutes. Extracellular fluid formula: 100mM glucose, 10mM CaCl 2 , 5mM MES (sodium α-sulfomethyl ester sulfonate), 0.8-1M sorbitol, and adjust the pH to 5.5-6.0 with Tris (tris...
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