Method for inducted rooting by taking coppice shoot of fir as explant

A technology for explants and tillering buds is applied in the field of inducing rooting and can solve the problems of high infection rate of explants, reduction of infection rate of explants and browning rate and the like

Inactive Publication Date: 2019-03-19
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the main problems in tissue culture of Chinese fir include: (1) Differences in genotypes lead to different requirements of Chinese fir for tissue culture, such as explant selection, basic medium, hormone ratio, etc., so it is necessary to establish a system for different clones (2) The problem of high infection rate of Chinese fir explants still exists, and it is urgent to screen out the most suitable disinfection and sterilization methods for different clone materials, so as to reduce the infection rate and browning of explants. (3) The multiplying multiples, rooting rate and the quality of proliferating seedlings and roots of Chinese fir explants need to be further improved

Method used

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  • Method for inducted rooting by taking coppice shoot of fir as explant
  • Method for inducted rooting by taking coppice shoot of fir as explant
  • Method for inducted rooting by taking coppice shoot of fir as explant

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Experimental program
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Effect test

Embodiment 1

[0038] The tiller buds of C54 and C64 clones were used as fir explants, and the materials were collected in early spring. The sterilization method used was: soaking in 75% alcohol for 30s+1% mercury chloride for 9 minutes, shaking for 1 minute, and then sterilized for later use.

[0039] Induction medium formula: 1 / 2MS, DCR, 3 / 4MS; 6-BA: 0.6mg / L, 0.8mg / L, 1.0mg / L; IAA: 0.1mg / L, 0.2mg / L, 0.3mg / L ; Sucrose 30g, carrageenan 8.5g, activated carbon 1.0g, pH 5.8. There are 9 treatments in total, 30 bottles are inoculated for each treatment, and 1 explant is inoculated in each bottle, the temperature is 25±2°C; the light is 12h.d-15h.d; the light intensity is 1500-2000lx. Data statistics were performed 30 days after induction. The formulations of each induction medium are shown in Table 2.

[0040] Table 2 Formula table of different induction medium

[0041]

[0042]

[0043] Induction of adventitious buds: The meristematic ability of the shoot tip is obviously stronger than...

Embodiment 2

[0058] The establishment of embodiment 2 Chinese fir subculture propagation system

[0059] Materials: C54 and C64 clone tissue culture seedlings cultivated in the induction test stage.

[0060] Subculture proliferation test with medium: MS, 1 / 2MS, DCR; 6-BA: 0.3mg / L, 0.6mg / L, 1.0mg / L; NAA: 0mg / L, 0.1mg / L, 0.2mg / L ; IBA: 0.1mg / L, 0.2mg / L, 0.3mg / L; sucrose 30g / L, carrageenan 8.5g / L, activated carbon 1.0g / L, pH=5.8; a total of 9 treatments, each treatment inoculated 20 Each bottle was inoculated with 4 to 6 explants; after 40 days of culture, the proliferation coefficient, germination number and growth of the explants were counted. The formulations of each subculture proliferation medium are shown in Table 7.

[0061] Table 7 Different Proliferation Medium Formulation Table

[0062] serial number

culture medium

6-BA(mg / L)

NAA(mg / L)

IBA(mg / L)

1

1 / 2MS

0.3

0

0.1

2

1 / 2MS

0.6

0.1

0.2

3

1 / 2MS

1.0

0.2

...

Embodiment 3

[0074] The establishment of rooting system in the Chinese fir bottle of embodiment 3

[0075] (1) Effect of basic medium on rooting induction

[0076] C54 clone tissue culture seedlings were used as rooting materials, basic medium 1 / 4DCR, 1 / 4MS, DCR, 0.8mg / L NAA, 20g / L sucrose, and carrageenan were adjusted according to the hardness of the medium. The formulas of the three rooting media are: ①1 / 4DCR+0.8mg / L NAA+20g / L sucrose+8.7g carrageenan; ②1 / 4MS+0.8mg / L NAA+20g / L sucrose+9.0g carrageenan; ③DCR+0.8mg / L NAA+20g / L sucrose+8.5g carrageenan.

[0077] During the culture process of induced rooting of Chinese fir tissue culture seedlings, it was observed that the adventitious roots would undergo the following morphological changes. Such as Figure 4 As shown, in the first 15 days, the tissue cultured seedlings grew well, and there was no adventitious root output; on the 15th day, the base of the tissue cultured seedlings expanded to form callus; There are adventitious roots, w...

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Abstract

The invention discloses a method for inducted rooting by taking a coppice shoot of a fir as an explant. The method comprises the following steps: 1) explant preparation and disinfection; 2) adventitious shoot induction; 3) adventitious shoot multiplication; and 4) adventitious shoot rooting. Proved by tests, the method for inducted rooting by taking the coppice shoot of the fir as the explant hasthe characteristics that the combined treatment of alcohol and mercury bichloride is more suitable for the disinfection of the fir explant, and the optimal disinfection way is that 75% alcohol is disinfected for 20-30 s and 0.1% mercury bichloride is disinfected for 8-10 min; the rooting effect and the tissue culture seedling survival rate on a DCR are obviously higher than those of an existing common 1/4 MS culture medium; an optimal multiplying culture medium beneficial to increment of multiplication times of each clone is DCR+0.6 mg/L6-BA+0.3 mg/LIBA; and a relatively proper method is provided for induced rooting of each fir clone individual.

Description

technical field [0001] The invention belongs to the technical field of plant breeding, and in particular relates to a method for inducing rooting by using fir tiller buds as explants. Background technique [0002] The breeding methods of Chinese fir include hybridization, cutting, grafting and tissue culture. Seed breeding is completed through male and female pollination, and there are often large and small years, and the breeding offspring will continue to undergo genetic recombination, which cannot maintain the stable transmission of the excellent genetic traits of the mother. This problem of separation of offspring can be solved by asexual reproduction. However, with the continuous expansion of the scale and management level of Chinese fir afforestation, cuttings and grafting are difficult to meet this huge demand. Therefore, tissue culture technology has begun to become an important technical means for the improvement and cultivation of Chinese fir clones, and its seedli...

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 边黎明杨坤施季森杨立伟李涛李若冰胡译胡俊渊季波刚
Owner NANJING FORESTRY UNIV
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