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A kind of adventitious bud induction and sub-proliferation culture method of A. chinensis

A technology of subculture propagation and culture method, which is applied in the field of adventitious bud induction and subculture propagation of Cyclocarya paliurus, can solve the problems of poor repeatability, poor callus growth state, and low differentiation rate of callus adventitious buds. Strong growth, reduced pollution rate and death rate, and increased induction rate

Active Publication Date: 2020-10-16
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing formulations generally have problems such as poor reproducibility, poor growth of adventitious buds and callus, and low callus differentiation rate.

Method used

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  • A kind of adventitious bud induction and sub-proliferation culture method of A. chinensis
  • A kind of adventitious bud induction and sub-proliferation culture method of A. chinensis
  • A kind of adventitious bud induction and sub-proliferation culture method of A. chinensis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Alcohol and sodium hypochlorite were selected to study the disinfection treatment of Cyclocarya paliurus, and the disinfection time of 75% alcohol: 25s, 30s. 10% sodium hypochlorite treatment time: 3min, 3.5min, 4min. The experimental design is detailed in Table 1. After trimming and rinsing, the stem segments are placed on an ultra-clean workbench, further disinfected with ethanol and sodium hypochlorite, and rinsed 4-5 times with sterile water after the disinfection is completed. Cut off the sterilized stem section with buds on the sterilized filter paper and cut off the part oxidized by the disinfectant and place it flat in the culture medium. After 10 days, observe the pollution of the stem section after inoculation, and observe the pollution characteristics of the explants to further judge Causes of pollution, and statistical pollution rate and death rate, the results are shown in Table 2.

[0033] It can be seen from Table 2 that for the pollution and death of e...

Embodiment 2

[0040] In order to study the effect of basic medium on the induction and proliferation of adventitious buds of Cyclocarya paliurus, five kinds of basic medium were set up in this experiment: MS, WPM, B5, DKW, 1 / 2MS (excluding macroelements and CaCl 2 1 / 2 of the full amount, and the rest are the full amount). The experiment was carried out by a single-factor randomized block experiment. The sterilized stems of Cyclocarya paliurus were placed on sterilized filter paper (11×11cm) to blot the water, and the oxidized parts at both ends of the stems were removed with sterilized scissors. and some petioles at the leaf axils, the pruned stems were inoculated in the culture medium, and the same hormone combination was added to the basic medium respectively. Part; the incision is inclined to increase the contact area between the cut surface and the medium; when the trimmed stem section is inserted into the high-temperature sterilized medium vertically, the insertion depth of the stem se...

Embodiment 3

[0047] In order to induce the first generation of adventitious buds from the stem segment of Cyclocarya paliurus, this experiment added auxomodulin 6-BA to the WPM basic medium, and mainly studied the effect of 6-BA concentration on the induction of adventitious buds in Cyclocarya paliurus. Table 4 below. The explants are selected from the sterilized, sterilized, and pruned stem segments in the previous stage, and are vertically inserted into the adventitious bud induction medium. The insertion depth accounts for about 1 / 4 of the length of the stem segment, and one stem segment is inserted into each bottle of medium , 20 bottles were inoculated at a time, and repeated 3 times. During this process, the environmental conditions in the cultivation room were controlled, and the contaminated materials and culture medium were cleaned up in time. After 20-25 days of cultivation, the induction of adventitious buds was counted, and the induction rate of clustered buds, the number of ne...

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Abstract

The invention discloses a method for inducing and multiplying adventitious buds of Cyclocarya paliurus, belonging to the technical field of plant tissue culture. Select the young shoots of Cyclocarya paliurus as explants, before the explants are induced and value-added culture, the explants are first sterilized and disinfected; the sterilized treatment adopts alcohol treatment for 25-30s, sodium hypochlorite treatment for 3-4min; Adventitious bud induction medium formula: WPM+6‑BA+sucrose+agar powder, and adjust pH to 5.75‑5.85; Cyclocarya adventitious bud proliferation medium formula: WPM+IBA+sucrose+agar powder. The present invention obtains a large amount of adventitious bud materials with strong growth, stretched leaves and high growth in a relatively short period of time by optimizing materials, sterilizing treatment and cultivating formula, laying the foundation for establishing an efficient in vitro rapid propagation system of Cyclocarya paliurus, and also It is of great significance to the protection, development and utilization of Cyclocarya paliurus germplasm resources.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture. In particular, it relates to a method for inducing and subcultured adventitious buds of Cyclocarya paliurus. Background technique [0002] Cyclocarya paliurus (Batal.) Iljinskaja is a plant of the genus Cyclocarya in the family Juglandaceae, widely distributed in the south of the Yangtze River at an altitude of 420-5200m (Flora of China, 1979). The active substances contained in its bark and leaves have the functions of clearing away heat and detoxification, relieving pain and reducing inflammation (China Traditional Chinese Medicine Resources Collection, 1994). The latest research shows that its unique secondary metabolites have good effects on blood lipid (WU ZF et al., 2017), blood lipid (Jiang et al., 2015; Ma et al., 2015), blood sugar (Huang Weilin et al., 2017) Wu Fan et al., 2014), enhancing immunity (Ren Wenkang et al., 2016), anti-oxidation (Wang et al., 2013; Tang et al....

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 洑香香成圆尚旭岚张洋方升佐杨万霞
Owner NANJING FORESTRY UNIV
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