Cyclocarya paliurus callus and suspension culture method for callus particles
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A technology of callus and suspension culture, applied in the field of plant tissue culture, can solve the problems of plant cell growth limitation and limited dissolved oxygen, and achieve the effects of increasing growth rate and strongly inhibiting enzyme activity.
Inactive Publication Date: 2012-06-13
JIAXING ZHEHUA AGRI TECH DEVCO
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[0009] Compared with solid culture, liquid suspension culture has good mixing, flow performance and higher mass trans
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Embodiment 1
[0030] Embodiment 1, construct a kind of cell suspension culture system for cultivating Cyclocarya paliurus callus, its steps are as follows:
[0031] 1 Take the young leaves of Cyclocarya paliurus as explants, wash and soak with detergent for 8 minutes, and rinse with running water for 1 hour. Under sterile conditions, soak in 75% alcohol for 20 seconds, take out, soak in 0.1% mercury liter for 15 minutes, wash with clean water 8 times, cut into 0.3-0.5cm 2 The organization block is spared.
[0032] 2 Inoculate the sterilized Cyclocarya paliurus explants on the callus induction medium under aseptic conditions, the medium is MS+1.0mg / ml6-BA+0.4mg / mlNAA, 30g / L sucrose, 5g / L agar , 24°C, 8 hours a day and 2000 Lux light for two weeks. The loose callus with good growth condition was selected to be cut and expanded by spin bottle, and continued to be cultivated for two weeks.
[0033] 3 Select five pieces of Cyclocarya paliurus callus with good growth condition ...
Embodiment 2
[0036] 1 Take the young leaves of Cyclocarya paliurus as explants, wash and soak with detergent for 8 minutes, and rinse with running water for 1 hour. Under sterile conditions, soak in 75% alcohol for 20 seconds, take it out, soak in 8% sodium hypochlorite for 15 minutes, wash with water 8 times, cut into 0.3-0.5cm 2 The organization block is spared.
[0037] 2 Inoculate the sterilized Cyclocarya paliurana explants on the callus induction medium under aseptic conditions, the medium is MS+2.0mg / ml6-BA+0.10mg / mlNAA, 30g / L sucrose, 5g / L agar , 24°C, 8 hours a day and 2000 Lux light for two weeks. The loose callus with good growth condition was selected to be cut and expanded by spin bottle, and continued to be cultivated for two weeks.
[0038] 3 Select five pieces of Cyclocarya paliurus callus with good growth condition and loose structure, each piece is about 1cm 2 , crushed with a scalpel and tweezers, inoculated in an Erlenmeyer flask filled with 200ml liquid ...
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Abstract
The invention provides a Cyclocarya paliurus callus and a suspension culture method for callus particles. The method mainly comprises the following steps: 1) selection and aseptic processing of plants, wherein, healthy Cyclocarya paliurus plants at an age of 3 to 7 months and 9 to 11 months are selected, annual twigs and tender leaves are cleaned, immersed in a washing agent for 5 to 20 min and flushed with clear water for 0.5 to 2 h, and under an aseptic condition, cleaned twigs and tender leaves are disinfected with alcohol having a concentration of 70 to 80% for 5 to 30 s and then are sterilized by using mercuric chloride having a concentration of 0.1% for 5 to 20 min or by using sodium hypochlorite having a concentration of 2 to 12% for 5 to 20 min; 2) induction culture of callus; 3) suspension culture of callus particles; and 4) amplification culture of callus. According to the invention, the growth rate of Cyclocarya paliurus callus can be greatly improved, and during a same period of time, when increment of callus on a solid medium reaches 721%, increment of callus obtained through fluid suspension culture reaches 1292%; according to results of experiments on inhibition of activity of alpha-glucosidase, in the process of fluid suspension culture, secondary metabolites are secreted to the exterior of cells, and callus cells and a culture solution both contain an alpha-glucosidase activity inhibitor.
Description
technical field [0001] The invention relates to a method for suspension culture of pacarya paliurus callus and callus particles, and belongs to the technical field of plant tissue culture. Background technique [0002] Cyclocarya papaya ( Cyclocarya Paliurus (Batal.) Iljinskaja is a Dicotyledoneae Juglandaceae plant. It is a rare single-species plant unique to my country and one of the endangered plants under national key protection. An economic tree species. [0003] It has been found through research that Cyclocarya paliurus contains various macro and trace elements necessary for the human body such as manganese, iron, copper, zinc, selenium, chromium, alum, germanium, etc., and is rich in triterpenoids, steroidal compounds, saponins, Compounds such as coumarin and flavonoids. These compounds can effectively reduce triglyceride and cholesterol, so that Cyclocarya paliurus has the functions of lowering blood pressure, blood sugar and blood fat, and has unique effe...
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