Long-term multiplication and storage method for embryonic callus of sweet potatoes

A technology of embryogenic callus and sweet potato, which is applied in the field of plant tissue culture, can solve the problems of short embryogenic retention time of sweet potato embryogenic callus, small number of embryogenic callus expansion, loss of viability and space, etc., to achieve guaranteed Mass proliferation, great application value, and long storage time

Inactive Publication Date: 2012-10-24
JIANGSU XUZHOU SWEET POTATO RES CENT
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AI Technical Summary

Problems solved by technology

The main problems existing in the current sweet potato embryogenic callus culture are: (1) Utilizing the current embryogenic callus suspension liquid proliferation technology, the embryogenic retention time of sweet potato embryogenic callus is short
(2) Embryogenic callus and non-embryogenic callus coexist, and are easily competed by non-embryogenic callus with fast value-added ability and lose viability and space and regeneration ability
(3) Embryogenic callus easily differentiates to form embryoid bodies, loses proliferation ability, and loses embryogenicity
(4) The selection of explants is limited by the growth period of sweet potato, and it is necessary to induce embryogenic callus in spring every year. The production cost is high, and the number of embryogenic callus propagation is small, which cannot meet the long-term requirements of the high-efficiency genetic transformation system. provided needs

Method used

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Embodiment Construction

[0015] The present invention will be described in detail below in conjunction with examples.

[0016] 1) Induction of embryogenic callus: Disinfect the shoot tips of sweet potato variety Xu 55-2 with 70% alcohol for 30 seconds, and immediately transfer them to 0.1% HgCl 2 Sterilize in solution for 3 minutes and wash 4-5 times with sterile water. Under the dissecting microscope, cut the shoot tip tissue (with 1-2 leaf primordia) of about 0.5mm in length, and culture the shoot tip tissue on the 2.0mg / L 2,4-D MS solid medium, and the culture condition is 28 °C, dark. Induce embryogenic callus.

[0017] 2) The induced embryogenic callus was cut into small pieces, and transferred into 2.0 mg / L 2,4-D MS solid medium, and the culture condition was 22°C, dark. After 45 days, under the Leica EZ4D microscopic camera system, select the spherical embryos and heart-shaped embryos with bright yellow color and dense granules, and transfer them into fresh 2.0mg / L2, 4-D MS solid medium, and...

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Abstract

The invention discloses a long-term multiplication and storage method for embryonic callus of sweet potatoes. The method comprises the processes of selection of successive embryonic callus, successive multiplication of the embryonic callus, and induction of the embryonic callus to mature embryo regeneration. The method has the advantages that: the process flow is simple, the multiplication rate of the embryonic callus is high, and the successive time is long; the technical problem that the long-term utilization of the embryonic callus is limited because the materials are limited by seasons and the embryonic retaining time is short and the like in the sweet potato tissue culture process is solved; a good receptor material is provided for the transgene technology of the sweet potatoes; and a large amount of efficient regenerated material is provided for annually producing the tissue culture seedlings of the sweet potatoes.

Description

technical field [0001] The invention relates to a plant tissue culture method, in particular to a method for sweet potato embryogenic callus to achieve multiplication and long-term subculture preservation through plant tissue culture and microscopic monitoring technology. Background technique [0002] Sweet potato (Ipomoea batatas (L.) Lam.) is an important food, feed, industrial raw material and new energy crop in the world. It is widely cultivated in tropical, subtropical and temperate regions such as Asia, Africa and Latin America. Sweet potato ranks seventh in the world's food production. In my country's grain production, it ranks fourth after rice, wheat and corn, and has important economic value and strategic position. At present, the perennial planting area of ​​sweet potato in my country is 5-6×10 6 hm 2 , accounting for more than 60% of the world's total area, and the annual total output accounts for more than 80% of the world's total output. [0003] Due to the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 王欣李强马代夫李秀英谢逸萍
Owner JIANGSU XUZHOU SWEET POTATO RES CENT
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