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A kind of induction method and application of the callus of D.

A technology of hair fern and callus, which is applied in the biological field, can solve the problems of difficult realization and easy browning, and achieve the effect of reducing browning

Active Publication Date: 2018-01-23
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the study of D. citrifolia tissue culture, because the process of inducing sporophytes through tissue culture takes a long period, and wild D. Somatic callus induction is difficult to achieve, and browning is easy to occur during callus induction, so there is no report on the callus induction of D. to report

Method used

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  • A kind of induction method and application of the callus of D.
  • A kind of induction method and application of the callus of D.
  • A kind of induction method and application of the callus of D.

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The experimental method is as follows:

[0050] 1. Induction of sporophytes

[0051] Take the mature spores and place them in a 1.5mL centrifuge tube, drop them into sterile water, oscillate fully to make a suspension, centrifuge at 4000r / min for 1min, make all the spores precipitate, and discard the supernatant. Then drop about 1.0mL of 0.1% HgCl into the centrifuge tube 2 solution, sterilized for 1 min, rinsed with sterile water 4 to 5 times, and obtained a sterile spore suspension by the above centrifugation method.

[0052] Different culture methods are then used, including solid medium culture and liquid medium culture. Solid medium adopts 1 / 2MS, 1 / 4MS and improved Knop's formula (NaH 2 PO 4 0.25g, KNO 3 0.25g, MgSO 4 0.25g, Ca(NO 3 ) 2 1.0g, distilled water 1000mL), the agar concentration is 0.8%. The culture medium was divided into Erlenmeyer flasks and sterilized under high pressure (121°C, 20min) for later use. After the prothallus grew out of the ...

Embodiment 2

[0060] 1. Induction of sporophytes

[0061] Take the mature spores and place them in a 1.5mL centrifuge tube, drop them into sterile water, oscillate fully to make a suspension, centrifuge at 4000r / min for 1min, make all the spores precipitate, and discard the supernatant. Then drop about 1.0mL of 0.1% HgCl into the centrifuge tube 2 solution, sterilized for 1 min, rinsed with sterile water 4 to 5 times, and obtained a sterile spore suspension by the above centrifugation method.

[0062] Different culture methods are then used, including solid medium culture and liquid medium culture. Solid medium adopts 1 / 2MS, 1 / 4MS and improved Knop's formula (NaH 2 PO 4 0.25g, KNO 3 0.25g, MgSO 4 0.25g, Ca(NO 3 ) 2 1.0g, distilled water 1000mL), the agar concentration is 0.8%. The culture medium was divided into Erlenmeyer flasks and sterilized under high pressure (121°C, 20min) for later use. After the prothallus grew out of the sexual organs, they were washed with sterile wate...

Embodiment 3

[0067] The difference between this embodiment and embodiment 1 is:

[0068] 1. Induction of sporophytes: inoculate it in the medium of 1 / 4MS+3.0% sucrose+0.8% agar at the stage of heart-shaped prothallus; cultivate it for a period of time and inoculate it in 1 / 2MS+3.0% sucrose+0.8% agar culture medium;

[0069] 2. Induction of green globules (GGB)

[0070] The sporophyte shoot tips of Trichomonas elegans obtained as explants were inserted into 1 / 2MS+1.0mg / L6-BA+3.0mg / L NAA+2.0% sucrose+0.7%~0.8% agar under sterile conditions GGB was obtained from the induction medium of T.

[0071] 3. Callus induction and proliferation

[0072] The obtained Trichophylla GGB was cultured by inoculating 1 / 2MS+1.0mg / L 6-BA+1.0mg / L 2,4-D+2.0% sucrose+0.7%~0.8% agar under sterile conditions The callus is formed in the base, and then added to 1 / 2MS + 3% sucrose + 0.8% agar medium for differentiation culture, subcultured once every 3-4 weeks, and the pH value of the medium is 5.7-5.8.

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Abstract

The invention discloses an induction method and application of dryopteris fragrans callus tissues, and belongs to the technical field of biology. The method provided by the invention comprises the following steps: preparing dryopteris fragrans spores into a sterile spore suspension, inoculating the sterile spore suspension to improved Knop' s culture mediums, performing dark treatment, and after germination, performing culturing so as to obtain sporophytes; using stem tips and young and tender leaf bases of the obtained dryopteris fragrans sporophytes as explants, inoculating the explants to culture mediums in which 6-BA and NAA are added, and performing culturing so as to obtain green spherules; inoculating stem tips of the green spherules or the stem tips of the sporophytes into culture mediums containing 6-BA and 2,4-D, and performing proliferation and differentiation so as to obtain the callus tissues. The method disclosed by the invention lays the theory and practice foundation for establishing a rapid propagation system of dryopteris fragrans, so that wild resources of the dryopteris fragrans are protected, and the blank of performing callus tissue induction through stem tips and young and tender leaf bases in the tissue culture research of the dryopteris fragrans is filled.

Description

technical field [0001] The invention relates to a method for inducing a callus of D. chrysalis and its application, and belongs to the field of biotechnology. Background technique [0002] Dryopteris fragrans (L.) Schott is a plant of the genus Dryopteris fragrans (L.) Schott, which is mainly distributed in China, Japan, Korea and other places, with Heilongjiang Province of China as the distribution center, and a large distribution area in Wudalianchi area . Dermascens mainly grows in talc slopes and volcanic magma crevices in alpine regions. Wang Quanxi, Momose et al. have done a lot of research on the gametophyte development of Trichofernaceae, but there is no report on the method of the callus induction and plant regeneration of Trichophylla. Drichopteris is a kind of medicinal plant, which has a good curative effect on skin diseases and has a certain development prospect. However, wild Drichopteris resources are being seriously damaged because of its medicinal value. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/00A01H4/001
Inventor 常缨卜志刚陈玲玲佟伟霜黄庆阳陶文青
Owner NORTHEAST AGRICULTURAL UNIVERSITY