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Method for carrying out generation of hybridized liriodendron chinense somatic embryos by utilizing salicylic acid

A technology for hybridizing Liriodendron tulipifera and somatic embryos of Liriodendron tulipifera, applied in horticultural methods, botanical equipment and methods, applications, etc., to achieve the effects of promoting large-scale application, improving efficiency, and reducing deformity rates

Active Publication Date: 2017-05-24
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, there have been many studies on the resistance to salicylic acid, but few studies in other fields, especially the effects of promoting cell growth and differentiation and inducing somatic embryogenesis under the conditions of plant tissue or cell culture in vitro.

Method used

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  • Method for carrying out generation of hybridized liriodendron chinense somatic embryos by utilizing salicylic acid
  • Method for carrying out generation of hybridized liriodendron chinense somatic embryos by utilizing salicylic acid
  • Method for carrying out generation of hybridized liriodendron chinense somatic embryos by utilizing salicylic acid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1. Establishment of hybrid Liriodendron suspension system

[0049] The embryogenic callus was transferred to 250 mL of callus liquid medium for shaking culture, the culture environment was dark culture, 24 ° C, and the shaker speed was 96 rpm. Callus of each genotype was inoculated into 3 flasks, and the suspension cell culture was prepared according to the ratio of 1:9, and the volume of each Erlenmeyer flask was 50 mL after the suspension cells were mixed with the culture medium. The subculture period of the material is 7 days, and after 2 subcultures, the cells are sieved. 400-mesh single cells were shaken and cultured in a liquid-adjusted medium for 1 day, and the medium was 3 / 4MS+NAA0.2mg / L+KT0.5mg / L+BA0.2mg / L+ABA1.0mg / L+VC5.0mg / L , sugar 50g / L, hydrolyzed casein 0.5mg / L, culture environment is dark culture, 24°C, shaker speed is 96rpm.

[0050] 2. Induction of somatic embryos in liquid medium

[0051] The liquid suspension cells over-cultured for 2 days were ob...

Embodiment 2

[0065] 1) After excessive liquid suspension culture, observe the state of liquid cells of different genotypes under a microscope, such as figure 2 shown. The hybrid Liriodendron tulipifera suspension cells of genotype C138 were selected for culture. Use a pipette gun to draw 2 mL and spread it on SA somatic embryo induction medium of different concentrations (SA concentration is number 1: 0 mg / L, number 2: 0.01 mg / L, number 3: 0.05 mg / L, number 4: 0.25 mg / L, No. 5: 1.25mg / L), observed and counted the number of somatic embryo development under a microscope after 28 days, and counted the number of somatic embryo seedlings after 45 days, and the statistical results are as shown in Table 4.

[0066] Table 4 The induction of somatic embryos with genotype C138 on different media

[0067]

[0068] Note: + grows poorly, ++ grows generally; +++ grows well; ++++ grows better; +++++ grows best, the same below.

[0069] As can be seen from Table 4, adding different concentrations ...

Embodiment 3

[0092] The hybrid Liriodendron tulipifera callus was directly transferred to the solid medium supplemented with different concentrations of SA, and 5 cell blocks were connected to each dish, and 6 dishes were inoculated at each concentration. The primary culture environment was dark culture. After the callus grew out of the embryonic structure, the light environment was adjusted to light culture for 16 hours, dark culture for 8 hours, and the temperature was 24°C. The specific addition concentration is shown in Table 10.

[0093] Table 10 SA addition concentration

[0094]

[0095]

[0096] Among them, agar is 8.0g / L, hydrolyzed milk protein is 0.5g / L, activated carbon is 4.0g / L, sucrose is 40g / L, pH5.7-5.8.

[0097] The genotypes of C138, 243012, and 253010 hybrid Liriodendron callus were respectively transferred to solid medium supplemented with different concentrations of SA (Table 10). After 28 days of culture, no obvious somatic embryos appeared. After two subcult...

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Abstract

The invention discloses a method for carrying out generation of hybridized liriodendron chinense somatic embryos by utilizing salicylic acid. The method comprises the following steps: establishing a hybridized liriodendron chinense suspension system, obtaining liquid single cells, transiting and inducing the generation of the hybridized liriodendron chinense somatic embryos; in a process of inducing the generation of the hybridized liriodendron chinense somatic embryos, adding the salicylic acid and ABA (Abscisic Acid) into a culture medium at the same time or independently adding the salicylic acid. According to the method for carrying out the generation of the hybridized liriodendron chinense somatic embryos by utilizing the salicylic acid, disclosed by the invention, a plant growth regulator SA is added in a generation process of the hybridized liriodendron chinense somatic embryos, so that the inductivity of the somatic embryos and the synchronism of development are effectively improved; benefits of breeding hybridized liriodendron chinense somatic cell engineering germchits are improved.

Description

technical field [0001] The invention belongs to the technical field of tissue culture seedlings, and in particular relates to a method for utilizing salicylic acid (SA) to perform somatic embryogenesis of a hybrid tulip tree. Background technique [0002] In recent years, people have continuously increased the research on plant somatic embryogenesis, which has achieved rapid development and achieved great results in decades. Because the somatic embryos induced by somatic cells have the characteristics of large number, fast growth, complete structure, and high reproductive efficiency, somatic embryogenesis technology has become an effective means for rapid plant reproduction, which is conducive to the preservation of excellent forest species and shortens the time for cultivating excellent varieties. Time, speeding up the development of industrialization, etc. are of great significance in the development of forestry. [0003] Plant somatic embryogenesis is an effective way fo...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 施季森施帅男陈金慧成铁龙杨立明陈婷婷陆叶
Owner NANJING FORESTRY UNIV
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