Cyclocarya paliurus adventitious bud inducing and subculture multiplication culture method

A technique of sub-proliferation and cultivation method, which is applied in the field of adventitious bud induction and sub-proliferation cultivation of A. The effect of reducing pollution rate and mortality, increasing induction rate and growing robustness

Active Publication Date: 2019-09-03
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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  • Cyclocarya paliurus adventitious bud inducing and subculture multiplication culture method
  • Cyclocarya paliurus adventitious bud inducing and subculture multiplication culture method
  • Cyclocarya paliurus adventitious bud inducing and subculture multiplication culture method

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Experimental program
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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 cyclocarya paliurus adventitious bud inducing and subculture multiplication culture method, and belongs to the technical field of plant tissue culture. Young cyclocarya paliurus twigs serve as explants, and before being subjected to inducing and multiplication culture, the explants are sterilized; during sterilization, alcohol is adopted to treat the explants for 20-30 s,and sodium hypochlorite is adopted to treat the explants for 3-4 min; a culture medium for cyclocarya paliurus adventitious bud inducing is prepared from WPM, 6-BA, saccharose and agar powder, and pHis adjusted to be 5.75-5.85; a culture medium for cyclocarya paliurus adventitious bud multiplication culture is prepared from WPM, IBA, saccharose and agar powder. According to the cyclocarya paliurus adventitious bud inducing and subculture multiplication culture method, through material optimization, sterilization and the culture formulas, a large quantity of adventitious bud materials which are healthy and strong, have spread leaves and grow rapidly, are obtained within a short time, the foundation is laid for building an efficient cyclocarya paliurus in-vitro rapid propagation system, and the cyclocarya paliurus adventitious bud inducing and subculture multiplication culture method also has great significance of protecting, developing and utilizing 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 Applications(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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