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A method of tissue culture and rapid propagation of Alisma fern seedlings via the green spheroid approach

A green spheroid, tissue culture rapid propagation technology, applied in the field of rapid propagation of Alisma fern through tissue culture, can solve the problems that there is no such thing as Alisma fern, and achieve the effects of short breeding cycle, well-developed root system, and high reproductive efficiency

Active Publication Date: 2016-01-20
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant report on the tissue culture propagation technology of Alisma fern green spheroid pathway

Method used

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  • A method of tissue culture and rapid propagation of Alisma fern seedlings via the green spheroid approach
  • A method of tissue culture and rapid propagation of Alisma fern seedlings via the green spheroid approach

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 The inventive method

[0035] (1) Acquisition of aseptic explants of Alismatis fern

[0036] Inoculate the aseptic spores of Alismatis fern on 1 / 2MS + 30.0g / L sucrose + 7.0g / L agar, pH 5.80 culture medium, the culture conditions are: 20~25℃, the daily light time is 12 hours, the light intensity It is 2000lx, and after cultivating for 6-7 months, young and tender aseptic seedlings can be obtained, and the young and tender aseptic seedlings are used as Alisma fern aseptic explants.

[0037] The method for obtaining the aseptic spores of Erythronia fern is to place 10-15 mg of spores in a 1.5 ml centrifuge tube, pre-wash them three times with sterile water, centrifuge at 6000 r / min for 2 minutes, obtain the spore precipitate, add a volume fraction of 5% NaClO solution to kill Bacteria for 4 minutes, washed with sterile water for 3-5 times to obtain aseptic spores of Alismatis fern.

[0038] (2) Induction of green spheroids of Alisma fern

[0039] Inoculate ...

Embodiment 2

[0058] Embodiment 2 The inventive method

[0059] Embodiment 2 is the same as Embodiment 1 except that the following steps are different, and will not be repeated here.

[0060] (2) Induction of green spheroids of Alisma fern

[0061] Green spheroid induction medium is: 1 / 6MS+TDZ0.6mg / L+NAA0.2mg / L+hydrolyzed casein 600mg / L+sucrose 30.0g / L+agar 7.0g / L, pH 5.80.

[0062] After 2 weeks of cultivation, the shoot tip and petiole swollen base of the aseptic seedlings will grow into green spheroids, and the average induction rate can reach 95%.

[0063] (3) Proliferation of the green spheroids of Alismatis fern

[0064] Green spheroid proliferation medium is: 1 / 6MS+TDZ0.2mg / L+6-BA0.8mg / L+NAA0.2mg / L+hydrolyzed casein 600mg / L+sucrose 30.0g / L+agar 7.0g / L, pH is 5.80.

[0065] After 4 weeks of culture, the fresh weight of the green spheroids increased by 137.52±6.36mg, and the multiplication factor was about 12.5 times.

[0066] (4) Differentiation of the green spheroids of Alisma f...

Embodiment 3

[0076] Embodiment 3 is the same as Embodiment 1 except that the following steps are different, and will not be repeated here.

[0077] (2) Induction of green spheroids of Alisma fern

[0078] Green spheroid induction medium is 1 / 5MS+TDZ1.0mg / L+NAA0.3mg / L+hydrolyzed casein 800mg / L+sucrose 30.0g / L+agar concentration 7.0g / L, pH is 5.80.

[0079] After 2 weeks of cultivation, the shoot tips and petiole bases of sterile seedlings swelled to form green spheroids, and the average induction rate could reach 98.33%.

[0080] (3) Proliferation of the green spheroids of Alismatis fern

[0081] Green spheroid proliferation medium is: 1 / 5MS+TDZ0.3mg / L+6-BA0.5mg / L+NAA0.3mg / L+hydrolyzed casein 800mg / L+sucrose 30.0g / L+agar 7.0g / L, pH is 5.80.

[0082] After 4 weeks of culture, the fresh weight of the green spheroids increased by 120.63±7.82 mg, and the multiplication factor was about 11 times.

[0083] (4) Differentiation of the green spheroids of Alisma fern

[0084] The green spheroid ...

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Abstract

The invention discloses a tissue culture and rapid propagation method of hemionitis arifolia seedlings by virtue of a green spherical body manner. Mediums used in the tissue culture and rapid propagation method comprise an induction medium, a propagation medium and a differential medium, wherein the induction medium is prepared from 0.4-1.0mg / L 1 / 6MS-1 / 4MS and TDZ, 0.1-0.3mg / L NAA and 400-800mg / L casein hydrolysate; the propagation medium is prepared from 0.1-0.3mg / L 1 / 6MS-1 / 4MS and TDZ, 0.5-1.0mg / L 6-BA, 0.1-0.3mg / L NAA and 400-800mg / L casein hydrolysate; the differential medium is prepared from 1 / 4MS-1 / 2MS and 3-6g / L active carbon; cane sugar and agar are added into all the mediums; the pH value of each medium is 5.80. The induction rate of the green spherical body is greater than 88%; the proliferation time is 11-12.5; the differentiation rate is greater than 98%; 1.5 million to 2 million of seedlings can be propagated from one green spherical body in one year; the survival rate s greater than 95%.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a rapid propagation method of Alisma alismas through tissue culture through green spheroids. technical background [0002] Alisma fern (Hemionitis arifolia) belongs to the genus Alisma fern of the gymnosperm family, trade name: wish fern, wish leaf, heart leaf fern. There are 8 species of Alisma genus, and only 1 species of Alisma is produced in Asia. China is the northern limit of the distribution of this species, mainly distributed in Hainan and southern Yunnan. The leaves of Alisma fern are heart-shaped and unique in shape, and have good market application prospects. [0003] Under natural conditions, ferns mainly rely on spores for reproduction, but the cycle of spore reproduction is long, usually 6-18 months, or even longer. [0004] The fern green spheroid refers to a spheroid composed of numerous green particles induced by fern explants, which is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 余蓉培吴学尉李绅崇李涵田敏蒋亚莲卢珍红周旭红
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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