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

Active Publication Date: 2017-08-01
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The induction of cassava embryogenic callus is complicated
Induction of fragile embryonic tissue is highly influenced by genotype, occurs infrequently, and takes a long time

Method used

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  • Method for culturing friable embryogenic calluses of manihot esculenta

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a method for culturing friable embryogenic calluses of manihot esculenta. The method comprises the following steps: (1) inducing the friable embryogenic calluses; (2) regenerating the friable embryogenic calluses. The method for culturing the friable embryogenic calluses of the manihot esculenta has the advantage that friable embryogenic callus induction is short in generation time and high in generation rate; the manihot esculenta is preferably the conventional cultivated varieties such as Huanan 8# and Huanan 6#.

Description

technical field [0001] The invention belongs to the technical field of brittle embryogenic callus culture, and in particular relates to a method for cultivating cassava brittle embryogenic callus. Background technique [0002] Cassava (Manihot esculenta Crantz) is a plant belonging to the genus Cassava in the Euphorbiaceae family, and is an important food and energy plant. People usually use traditional breeding methods to improve its quality, but some traits, such as resistance to diseases and insect pests, and reduced cyanide content, can only be improved through genetic engineering. An effective regeneration system is a prerequisite for the successful implementation of genetic engineering. Fragile embryogenic callus is a tissue with a diameter of ≤1mm, which is easy to disperse and proliferates quickly in the liquid culture process, and most of the cells are totipotent, so high-quality embryogenic suspension cultures can be obtained. Because it can increase the probabil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/00
Inventor 李瑞梅郭建春姚远刘姣段瑞军符少萍
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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