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A medium for callus induction and sub-proliferation of C. chinensis and its culturing method

A technology of callus induction and subculture, which is applied in the field of plant tissue culture, can solve the problems of scattered distribution of natural resources and obvious differences in component content, and achieve the effects of easy handling and management, good response effect, and increased proliferation rate

Active Publication Date: 2020-10-30
NANJING FORESTRY UNIV
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  • Abstract
  • Description
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Problems solved by technology

The medicinal components of Cyclocarya paliurus are mainly extracted from the leaves. However, the distribution of natural resources is scattered, and the content of components between provenances and individuals varies significantly. Therefore, in order to effectively protect the germplasm resources and realize the industrial production of secondary metabolites, organizations The culture technique that promotes the biosynthesis of secondary metabolites at the tissue cell level is of special theoretical and practical significance

Method used

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  • A medium for callus induction and sub-proliferation of C. chinensis and its culturing method
  • A medium for callus induction and sub-proliferation of C. chinensis and its culturing method
  • A medium for callus induction and sub-proliferation of C. chinensis and its culturing method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, the selection of explant and the screening of disinfection time

[0046] Selection of explants: In 2017, we collected healthy and disease-free vegetative branches from the Qingqian Liu Nursery of the Baima Base of Nanjing Forestry University and the Greenhouse on the 10th Floor of the Biotechnology Building of Nanjing Forestry University, and cut long branches from the top 1 and 2 nodes. About 3-4cm semi-lignified stem section with buds, retaining 1 axillary bud.

[0047] Pruning and cleaning of explants: trim the leaves on the stems, keep the petioles and 1 / 5 of the leaves, and rinse with tap water for 1 to 2 hours.

[0048] In order to screen out the best treatment combination of disinfection time for the stems of Cyclocarya paliurus, two disinfectants, alcohol and sodium hypochlorite, were selected to study the disinfection treatment of Cyclocarya paliurus, and the disinfection time of 75% alcohol was selected at two levels: 25s and 30s. 10% sodium hyp...

Embodiment 2

[0057] Embodiment 2, anti-browning agent concentration screening

[0058] In the experiment, it was found that the stem section was browned after inoculation. In order to study the effect of the anti-browning agent PVP (polyvinylpyrrolidone) on the browning degree of the callus of Cyclocarya paliurus, the concentration of PVP was set at 4 levels in this experiment: 0 , 100mg·L -1 , 150mg·L -1 , 200mg·L -1 . The experiment was carried out with a single factor randomized block experiment. The sterilized stems of Cyclocarya paliurus were placed on sterilized filter paper (11×11cm) to dry, and the ends of the stems and the axils of the leaves were removed with sterilized scissors. Petiole, put the cut stem segment flat on the sterilized callus induction medium (the medium formula is WPM+1.5mg L -1 6-BA+0.3mg·L -1 IBA), add different concentrations of PVP respectively, the depth of the stem section is 1 / 4 of the length of the stem section, put 2 to 3 stem sections in each bot...

Embodiment 3

[0063] Embodiment 3, sterile anti-browning system callus induction medium formulation screening

[0064] In order to induce callus from the stems of Cyclocarya paliurus, 200mg·L -1 PVP, 6-BA and NAA, add 200mg·L in WPM basic medium -1 PVP, 6-BA and IBA two hormones. The effect of growth regulator concentration on callus induction was mainly studied, and the combination of hormone concentration and treatment is shown in Table 4 below. The explants were selected from the stem section of the sterile culture system constructed in the previous stage, trimmed and placed flat in the callus induction medium. The depth of the flat part accounted for 1 / 4 of the length of the stem section. ~3 stem segments, inoculate 20 bottles at a time, and repeat 3 times. During this process, control the environmental conditions in the cultivation room, and clean up the contaminated and browned materials and culture medium in time. After 20 days of culture, the callus induction was counted.

[0...

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Abstract

The invention discloses a culture medium for Cyclocarya paliurus callus induction and subculture proliferation, and also discloses a culture method based on Cyclocarya paliurus callus induction and subculture proliferation medium, including selection of explants, selection of explants Sterilization, establishment of sterile anti-browning system and proliferation culture. The invention can effectively improve the anti-browning ability of Cyclocarya paliurus callus, obtain a large amount of callus without browning and fast growth in a relatively short period of time, and solve the problem of shortage of Cyclocarya paliurus resources by means of biotechnology. In the later stage of industrialization, the secondary metabolites such as triterpenoids, flavonoids, and polysaccharides of Cyclocarya paliurus were extracted to lay the foundation.

Description

technical field [0001] The invention relates to a papaya paliurus callus induction and subculture culture medium and a culture method thereof, belonging to the technical field of plant tissue culture. Background technique [0002] Cyclocarya paliurus (Batal.) Iljinskaja belongs to Dicotyledoneae (Dicotyledoneae), Juglandaceae (Juglandaceae) Cyclocarya (Cyclocarya), is an important medicinal and edible homologous plant, and is a unique single plant in China. Plants of the genus (Flora of China, 1979), in recent years, because of the unique triterpene flavonoid polysaccharides contained in its leaves, it has been proved to have antihypertensive (Hou Xiaoli et al., 2014), hypolipidemic (Jiang et al, 2015; Ma et al , 2015), hypoglycemic (Yoshitomi et al, 2017; Huang Weilin et al, 2017), anti-oxidation (Wang et al, 2015; Xie et al, 2015) and other physiological functions, and has the ability to improve kidney function, regulate blood sugar disorders, and relieve liver damage , I...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 成圆洑香香张洋尚旭岚方升佐
Owner NANJING FORESTRY UNIV
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