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Light regulation and control method for tissue culture and multiplication of camptotheca acuminata decaisne calluses

A technology of callus and light regulation, applied in the field of plant cultivation, can solve problems such as mercury content, environmental pollution, and single light quality, and achieve the effects of increasing induction rate and proliferation rate, reducing production costs, and reducing environmental pollution

Inactive Publication Date: 2014-01-08
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, fluorescent lamps are the main light source for tissue culture, but they are high in energy consumption, short in life, contain mercury, are easy to cause environmental pollution, and have a single light quality, which cannot meet the different needs of plants in different growth stages of tissue culture.

Method used

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  • Light regulation and control method for tissue culture and multiplication of camptotheca acuminata decaisne calluses
  • Light regulation and control method for tissue culture and multiplication of camptotheca acuminata decaisne calluses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Take the young stems of Campyloditi radicis that are free from diseases and insect pests in the same year as explants. After picking and cleaning the stems, sterilize them in 75% ethanol solution for 1 min, put them into 0.1% mercuric chloride solution for further disinfection for 5 min, rinse them with sterile water for 3-4 times, dry them with sterile filter paper, and set aside.

[0017] Cut the stems into 2-3 mm small pieces, and inoculate them into the following four callus induction media respectively, inoculate 15 bottles of each medium, and inoculate 5 explants in each bottle, and transfer to Cultivate under light, culture temperature (25±2)°C, light intensity 1000-1500lx, light time 12h / day, light source is fluorescent lamp. The experiment was repeated 3 times.

[0018] ①MS+0.1mg / L KT+4mg / L2,4-D

[0019] ②MS+0.5mg / L KT+4mg / L2,4-D

[0020] ③MS+2mg / L6-BA+2mg / L NAA+2mg / L2,4-D

[0021] ④MS+0.5mg / L6-BA+2mg / L NAA

[0022] The agar of the above 4 kinds of media i...

Embodiment 2

[0025] Subculture the callus induced on No. ① and No. ② media respectively, and the subculture media used are:

[0026] ④MS+2.0mg / L6-BA+0.5mg / LNAA

[0027] ⑤MS+0.5mg / L KT+4.0mg / LNAA

[0028] (⑥B5+0.5mg / L KT+1.0mg / LNAA+0.5mg / L2, 4-D

[0029] The subculture conditions are culture temperature (25±2)°C, light intensity of 1000-15001x, light time of 12h / day, and fluorescent lamp as light source.

[0030] After 30 days of subculture, observe and count the subculture of induced callus on No. ① and No. ② media, and find that: the subculture of callus induced on No. ① media grows well on B5 (⑥ No.) media , more than 85% of the calli are loose in structure, easily scattered, light and transparent in color, and are good calli, while some of the calli subcultured on MS (4 and 5) medium have water stains on the surface Some are too compact and grow slowly, which is not conducive to the continued growth of callus; the subculture of callus induced on medium ② is not good, and it is found ...

Embodiment 3

[0033] The excellent callus subcultured on No. ⑥ medium was transferred and proliferated. The transfer medium was still No. ⑥, and the transfer container was a petri dish with a specification of 90*15mm. After the transfer, the petri dish was placed in a fluorescent lamp and Cultivate under 5 LED light sources with different light qualities, put 9 petri dishes under each light, and repeat 3 times. Adjust the LED light control software and the distance between the light source and the petri dish so that the amount of light is 105 μmol m -2 ·s -1 around, the photoperiod is 16h·d -1 , cultivated for 30 days. The relative humidity of the culture room is 70%±5%, and the temperature is (24±1)°C.

[0034] Wherein, the light source is an LED lamp purchased from Hangzhou Hanhui Photoelectric Technology Co., Ltd. See Table 1 for specific technical parameters.

[0035] Table 1 Main technical parameters of different light quality LED light sources

[0036]

[0037] After 30 days ...

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Abstract

The invention discloses a light regulation and control method for tissue culture and multiplication of camptotheca acuminata calluses, which sieves an optimal culture medium formula for establishing a camptotheca acuminate clone. A result shows that an optimal induction culture medium of stems contains MS, 0.1mg / L of KT and 4mg / L of 2,4-D; the optimal multiplication conditions are as follows: B5, 0.5mg / L of KT, 1.0mg / L of NAA (Naphthalene Acetic Acid) and 0.5mg / L of the 2,4-D; leaves represent poor inductivity. The calluses are multiplied under the conditions of red light, green light, blue light, red-blue light and white light; by analyzing the increment of calluses in unit, the red light and the green light have an acceleration effect on the growth of the calluses, the blue light has the inhibition effect on the growth of the calluses, and the effects of the white light and the red-blue light are not obvious; the acceleration effect of the green light is the most significant.

Description

technical field [0001] The invention belongs to the technical field of plant cultivation, and relates to a method for light regulation of camphor tree callus culture and proliferation. Background technique [0002] Campototheca acuminate Decaisne is a deciduous tree of the Davidiaceae family, a unique tree species in my country. In 1966, Dr. Monroe E.WALL of the United States isolated camptothecin (Compothecine, referred to as CPT) from the secondary metabolites of camptotheca. In 1985, Hisang et al. found that CPT can block the synthesis of topoisomerase I and have anticancer effects. At present, a variety of camptothecin derivatives synthesized with CPT as a precursor have been used in the clinical treatment of ovarian cancer, colon cancer, etc. In addition, CPT also has a strong inhibitory effect on HIV virus. With the continuous excavation and verification of CPT's anti-tumor and anti-virus functions, the global demand for CPT has increased rapidly. Since the 1990s, c...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 黄华宏林二培楼雄珍童再康李许可
Owner ZHEJIANG FORESTRY UNIVERSITY
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