Culture method capable of promoting cyclocarya paliurus callus growth and secondary metabolite accumulation

A technology for secondary metabolites and callus, applied in the field of plant tissue culture, can solve problems such as accumulation of secondary metabolites that are unfavorable for callus growth, and achieve the promotion of tight texture, rapid subculture, and low overall pollution rate. , the effect of inhibiting the production of phenolic substances

Active Publication Date: 2019-11-12
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the callus of Cyclocarya paliurus tends to produce phenolic substances during the culture process, which is not conducive to the growth of callus and the accumulation of secondary metabolites

Method used

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  • Culture method capable of promoting cyclocarya paliurus callus growth and secondary metabolite accumulation
  • Culture method capable of promoting cyclocarya paliurus callus growth and secondary metabolite accumulation
  • Culture method capable of promoting cyclocarya paliurus callus growth and secondary metabolite accumulation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, disinfection method screening

[0038] Selection of stem segments: Cut semi-lignified bud stem segments with a length of about 3 to 4 cm from the top 1 and 2 nodes of the three-year-old stubble seedlings cultivated in the greenhouse on the 10th floor of the Biotechnology Building of Nanjing Forestry University in 2018, and keep 1 Axillary buds.

[0039]Material pretreatment: Trim off the leaves on the stems, rinse with tap water for 1-2 hours.

[0040] In order to screen out the best treatment combination of disinfection time for the stems of Cyclocarya paliurus, two disinfectants of 0.1% mercury liter and 5% sodium hypochlorite were selected for research on the disinfection treatment of Cyclocarya paliurus, and the disinfection of 0.1% mercury liter and 5% sodium hypochlorite 5 levels of time are used: 3, 5, 7, 9, 11 minutes. The experimental design is detailed in Table 1. After trimming and rinsing, the stem segments were placed on a clean workbench, ...

Embodiment 2

[0047] Embodiment 2, callus induction formula screening

[0048] In order to induce callus from stems and leaves of Cyclocarya paliurus, 6-BA (0.5, 1.0, 1.5 mg·L -1 )+NAA (0.05, 0.1mg·L -1 ) hormone combination; with WPM as the basic medium, design 6-BA (0.5, 1.0mg·L -1 )+IBA (0.05, 0.1mg·L -1 ) hormone combination is used for the callus induction of stem segment and blade, and the specific operations are shown in Table 2 below. Each test tube was inoculated with one material, 15 tubes were treated each time, and repeated three times. Observe every 10 days, and calculate the callus induction rate after 30 days of culture.

[0049] Table 2 Screening of callus induction formula of Cyclocarya paliurus

[0050] Table2 Selection of callus induction formula for C. paliurus

[0051]

[0052] Note: + indicates that the number of callus is average, ++ indicates that the number of callus is large, the same below.

[0053] Comparing the effects of different hormone combinations...

Embodiment 3

[0057] Embodiment 3, callus proliferation formula screening

[0058] Proliferation culture: using WPM as the basic medium, according to the formula results in Table 2, three formulas were screened out as callus proliferation formulas, and the specific formulas are shown in Table 3. Each test tube was inoculated with one material, 15 bottles were treated each time, and repeated three times. Observed every 10 days, counted callus increment and growth multiple after 30 days of culture, and counted the growth status of callus after 7 passages. (Growth factor = (weight after subculture once - weight before subculture) / weight before subculture)

[0059] Table 3 Screening of callus proliferation formula of Cyclocarya paliurus

[0060] Table3 Selection of proliferation coefficient formula for callus of C. paliurus

[0061]

[0062] Table 4 Callus growth status of Cyclocarya paliurus after 7 subcultures

[0063] Table4 Growth state of seven successive generations for callus of C...

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Abstract

The invention discloses a culture method capable of promoting cyclocarya paliurus callus growth and secondary metabolite accumulation. The culture method includes: taking materials, and disinfecting;selecting semi-lignified stems with buds and top-end leaflets of cyclocarya paliurus, and disinfecting; performing callus induced culture: culturing stems and leaves in an induced culture medium to generate callus, wherein culture conditions are continuous dark treatment, and culture temperature is set at 20 DEG C; performing callus multiplication culture: transferring the callus into a transgenerational multiplication culture medium for transgenerational multiplication culture, and using a sealing film to sea, and putting into a culture chamber for culture, wherein culture conditions are continuous dark treatment, and culture temperature is set at 20 DEG C. Induction and multiplication culture of the callus are performed at temperature of 20 DEG C, so that generation of phenolic substances is inhibited while normal growth of cyclocarya paliurus is ensured, and callus growth and secondary metabolite accumulation are promoted at low temperature.

Description

technical field [0001] The invention relates to a culture method for promoting the growth of Cyclocarya paliurus callus and the accumulation of secondary metabolites, and belongs to the technical field of plant tissue culture. Background technique [0002] Cyclocarya paliurus (Batal.) Iljinskaja belongs to Dicotyledoneae (Dicotyledoneae) Juglandaceae (Juglandaceae) Cyclocarya (Cyclocarya) plant, it has broad application prospects in ornamental, medicinal, and timber. In terms of viewing, the tree shape is beautiful and straight, and the fruit has disc-shaped fruit wings, which look like copper coins from a distance; the root system is developed and the root tiller is strong. It is an excellent landscape afforestation tree species. In terms of material use, its growth rate is fast, the thickness of annual rings is uniform, the wood strength is high, and it is wear-resistant. It is suitable for furniture, interior decoration and building materials. In terms of medicinal use, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 洑香香张洋程锦萍成圆尚旭岚方升佐
Owner NANJING FORESTRY UNIV
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