Method of somatic embryogenesis of cassava and rapid propagation of regenerated plant
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A technology for plant regeneration and embryogenesis, which is applied in the fields of botany equipment and methods, plant cells, and plant regeneration, and can solve problems such as no domestic reports.
Inactive Publication Date: 2009-01-21
朱文丽 +4
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[0010] (3) Aspect test-tube seedling hardening and transplanting, mastered the light and temperature conditions and the required time r
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Embodiment 1
[0024] Embodiment 1: adopt the stem segment that has lateral bud to carry out microbody rapid propagation
[0025] The test variety is Huanan 5;
[0026] Disinfection treatment of explants: cut cassava stems into 30cm long cuttings, insert them vertically in the river sand substrate for cultivation, and after the side buds germinate and grow into new branches, remove the leaves, soak in saturated soapy water for 20 minutes, and then rinse with tap water 20min. After drying, soak in 70% alcohol for 4 seconds on the ultra-clean workbench, then soak in 0.2% mercuric chloride solution for 6 minutes, and rinse with sterile water for 5 times. Finally it is cut into about 2.0cm long stem section with side buds as explants;
[0027] Initial culture: the explants were inoculated on the initial medium: MS+GA 0.02mg / L+NAA 0.02mg / L+sucrose 30g / L+carrageenan 5.6g / L (pH=5.8). Then placed at a temperature of 28°C, cultivated in light for 12 hours a day, with a light intensity of 2000 lx, an...
Embodiment 2
[0031] The somatic cell regeneration system of embodiment 2 cassava
[0032] The test variety is Huanan 8;
[0033] Disinfection treatment of explants: After removing the leaves growing on the new shoots, soak them in saturated soapy water for 20 minutes, then rinse them with tap water for 20 minutes. After drying, soak in 70% alcohol for 4 seconds on the ultra-clean workbench, then soak in 0.2% mercuric chloride solution for 8 minutes, and rinse with sterile water for 5 times. Finally it is cut into about 2.0cm long stem section with side buds as explants;
[0034] Initial culture: the explants were inoculated on the initial medium: MS+GA 0.02mg / L+NAA 0.02mg / L+sucrose 30g / L+carrageenan 5.6g / L (pH=5.8). Then placed at a temperature of 28°C, cultivated in light for 12 hours a day, with a light intensity of 2000 lx, and cultivated for 40 days;
[0035] Subculture: after culturing for 40 days, a seedling of about 7 cm grows on the explant, and the stem section with lateral bud...
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Abstract
The invention relates to a tissue culture propagating method of Manihot esculenta Crantz. As one of the global three-top tuber crops and the number one alimentarn crop in Africa, the Manihot esculenta Crantz is currently ranked as a vital biological energy crop in our country. Stem section vegetative cottage is commonly adopted for propagation, and the low propagation rate thereof is a restrictive factor for popularizing the new variety and shortening the breeding period. The method of the invention comprises the following steps: the Manihot esculenta Crantz stem section with stem apexes or lateral buds is taken as an explant, and the Manihot esculenta Crantz tissue culture plantlets are quickly obtained in virtue of the micro-propagation method; young leaves, stem apexes and axillary buds of the sterile plantlets obtained from micro-propagation are taken as explants, somatic embryo generation and plant regeneration are induced, thus the Manihot esculenta Crantz somatic cell regeneration system is established. By adopting the method, the Manihot esculenta Crantz has the advantages of high propagation rate, no genotype dependence, and the like; the method has high value on the quick propagation and large-scale production of the Manihot esculenta Crantz improved variety in a short term, and establishes a technical basis for the Manihot esculenta Crantz transgene and other breeding.
Description
1. Technical field [0001] The invention relates to a plant tissue culture method, in particular to a method for achieving rapid propagation and large-scale production of cassava varieties through biotechnology. 2. Background technology [0002] Technical practicality analysis: [0003] Cassava (Manihot esculenta Crantz) is a plant of Euphorbiaceae (Euphorbiaceae), the number of chromosomes is 2n=36, and it is a perennial shrub with a height of 1-5m. Cassava is the fourth largest tropical crop after rice, corn, and sorghum. Its roots are rich in starch and can be used not only as food, but also as industrial raw materials or feed. It is the main source of food for 600 million people in tropical and subtropical regions. Cassava originated in South America and was introduced to Africa, Asia and Oceania in the 16th to 18th centuries. Cassava is now widely distributed in tropical and some subtropical regions of the world. Cassava was introduced into my country by overseas Chin...
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