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Induction method and application of dryopteris fragrans callus tissues

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

Active Publication Date: 2016-07-20
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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  • Induction method and application of dryopteris fragrans callus tissues
  • Induction method and application of dryopteris fragrans callus tissues
  • Induction method and application of dryopteris fragrans callus tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The experimental method is as follows:

[0050] 1. Induction of sporophyte

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

[0052] Then different culture methods were adopted, including solid medium culture and liquid medium culture. Solid medium using 1 / 2MS, 1 / 4MS and modified 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 conical flasks, autoclaved (121 ℃, 20 min) for later use. Rinse with sterile water every 3 days after the proth...

Embodiment 2

[0060] 1. Induction of sporophyte

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

[0062] Then different culture methods were adopted, including solid medium culture and liquid medium culture. Solid medium using 1 / 2MS, 1 / 4MS and modified 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 conical flasks, autoclaved (121 ℃, 20 min) for later use. Rinse with sterile water every 3 days after the prothallus grows the sex organs to promote fertilizati...

Embodiment 3

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

[0068] 1. Induction of sporophyte: inoculate it in the medium of 1 / 4MS + 3.0% sucrose + 0.8% agar at the stage of heart-shaped prothallus for cultivation; cultivate for a period of time until the prothallium grows sexual organs, inoculate it in Culture in 1 / 2MS+3.0% sucrose+0.8% agar medium;

[0069] 2. Induction of Green Globules (GGB)

[0070] The obtained sporophyte shoot tips of Trichoderma serrata were used as explants, and were inserted into 1 / 2MS+1.0mg / L6-BA+3.0mg / LNAA+2.0% sucrose+0.7%~0.8% agar under sterile conditions. In the induction medium, GGB was obtained.

[0071] 3. Callus induction and proliferation

[0072] The obtained Pteridophyta GGB was added to the medium of 1 / 2MS+1.0mg / L6-BA+1.0mg / L2,4-D+2.0% sucrose+0.7%~0.8% agar under sterile conditions The callus is formed, and then added to 1 / 2MS+3% sucrose+0.8% agar medium for differentiation and culture, and subcultures once every 3-4 weeks...

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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 and application for inducing callus of Trichosanthes serrata, and belongs to the field of biotechnology. Background technique [0002] Dryopteris fragrans (L.) Schott is a genus of Pteridaceae, mainly distributed in China, Japan, Korea and other places, with Heilongjiang Province as the distribution center, and a large distribution area in Wudalianchi area. Pteridophyta grows mainly 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 Pteridaceae, but there is no report on the method of callus induction and plant regeneration of Pteridophyta. Pteridophyta is a medicinal plant, which has a good curative effect on skin diseases and has a certain development prospect, and the wild resources of P. Using fern spores for tissue culture can establish a rapid propagation system of ferns. In the research on tissue culture of Pte...

Claims

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Application Information

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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
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