Method for culturing friable embryogenic calluses of manihot esculenta
An embryogenic callus and culture method technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of low frequency of occurrence, long induction time, and great influence of brittle embryogenic tissue, and achieve the production of High rate and short induction time
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Embodiment 1
[0044] The culture method of the cassava brittle embryogenic callus provided by the present embodiment comprises the following steps:
[0045] (1) Induction of brittle embryogenic callus
[0046] (1.1) The purified cassava body embryo block is placed in the triangular flask that 300mmol / L mannitol is housed, and is sealed with air-permeable sealing film; Put the above-mentioned triangular flask into a vacuum pump, open the switch, close the intake valve, when the air pressure drops to Start timing at 0.08Mpa, and take it out after 15 minutes; take out the above-treated somatic embryos in a sterilized ultra-clean workbench, blot the water with sterilized filter paper, place it on FECIM for 20 days in dark culture, and induce primary brittle embryogenic callus;
[0047] (1.2) Under a stereo microscope, the primary brittle embryogenic callus was transferred to fresh FECIM for proliferation and expansion, and a large amount of brittle embryogenic callus was obtained once every 2 ...
Embodiment 2
[0061] The culture method of the cassava brittle embryogenic callus provided by the present embodiment comprises the following steps:
[0062] (1) Induction of brittle embryogenic callus
[0063] (1.1) The cassava body germ block of purification is placed in the Erlenmeyer flask that 300mmol / L mannitol is housed, seals with air-permeable sealing film;
[0064] Put the above-mentioned triangular bottle into the vacuum pump, turn on the switch, close the intake valve, start timing when the air pressure drops to 0.08Mpa, and take it out after 20min;
[0065] In a sterilized ultra-clean workbench, take out the treated somatic embryo blocks, blot the water with sterilized filter paper, place them on FECIM for 25 days in the dark, and induce the production of primary brittle embryogenic callus;
[0066] (1.2) Under a stereo microscope, the primary brittle embryogenic callus was transferred to fresh FECIM for proliferation and expansion, and a large amount of brittle embryogenic cal...
Embodiment 3
[0080] The culture method of the cassava brittle embryogenic callus provided by the present embodiment comprises the following steps:
[0081] (1) Induction of brittle embryogenic callus
[0082] (1) No. 8 cassava body embryo pieces in South China of purification are placed in the triangular flask that 250mmol / L mannitol is housed, seal with air-permeable sealing film;
[0083] (2) Put the above-mentioned triangular flask into the vacuum pump, turn on the switch, close the intake valve, and start timing when the air pressure drops to 0.08Mpa. After 20min, take it out;
[0084] (3) In a sterilized ultra-clean workbench, take out the treated somatic embryo blocks, blot the water with sterilized filter paper, place them on FECIM for 30 days in dark culture, and induce primary brittle embryogenic callus;
[0085] (4) Under a stereo microscope, the primary brittle embryogenic callus was transferred to fresh FECIM for proliferation and expansion, and a large amount of brittle embr...
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