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Ultralow-temperature storage and regenerative-culture method for embedding and drying of isolated stem tips of test-tube horseradish plantlets

A technology of cryopreservation and regeneration culture, which is applied in the field of germplasm preservation, and can solve the problems of low regeneration rate, none, and plant variation

Active Publication Date: 2012-10-03
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

It is known that there are two methods of cryopreservation of wasabi: vitrification method and embedding vitrification method, both of which have the problems of plant variation and low regeneration rate, and there is no research report on a more effective method

Method used

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  • Ultralow-temperature storage and regenerative-culture method for embedding and drying of isolated stem tips of test-tube horseradish plantlets

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Embodiment Construction

[0030] The specific implementation manners of the present invention will be further described below in conjunction with the accompanying drawings.

[0031] Such as figure 1 The shown wasabi test-tube plantlet isolated shoot tip embedding and drying method cryopreservation and its regeneration culture method include the following specific steps:

[0032] -Material propagation: culture sterile seedlings in MS + 0.1mg / L BA + 20g of 825 mg / L NH4NO3, 950mg / L KNO3, 220mg / L CaCl2 2H2O, 185mg / L MgSO4 7H2O, 85mg / L KH2PO4 / L sucrose + 7g / L agar medium, pH 5.8, temperature 20±2°C, light time 16h, subculture once every 35-40d. It was found in the experiment that the content of NH4NO3, KNO3, CaCl2·2H2O, MgSO4·7H2O, KH2PO4 in the MS medium was halved, and the growth and propagation effects of sterile seedlings were good.

[0033] - Stem tip stripping: under aseptic conditions, isolated shoot tips with a diameter of about 1 to 3 mm are cut from healthy sterile seedlings as materials.

[...

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Abstract

The invention discloses an ultralow-temperature storage and regenerative-culture method for embedding and drying of isolated stem tips of test-tube horseradish plantlets, and relates to the technical field of germplasm preservation. The ultralow-temperature storage and regenerative-culture method comprises the following steps of: propagating materials, stripping the stem tips, preculturing, embedding calcium alginate into balls, drying the balls with sterile air flow, putting the dried balls into a freezing pipe, putting the freezing pipe in liquid nitrogen for ultralow-temperature storage, thawing, and restoring the regenerative culture. After ultralow-temperature storage, the isolated stem tips of the test-tube horseradish plantlets are treated by thawing, and the seedlings are directly formed after the regenerative culture is restored, so that the hereditary variation is avoided. The ultralow-temperature storage and regenerative-culture method is long-term, safe, stable, reliable and simple, and also has the advantages that the germplasm resource of the horseradish is effectively stored, the restoration and the recovery growth condition of the regeneration seedlings is favorable and the plant regeneration rate is high.

Description

technical field [0001] The invention relates to the technical field of germplasm preservation, in particular to a cryopreservation method and a recultivation method of plant germplasm resources. Background technique [0002] In the preservation of germplasm resources, for the safety of stored germplasm, it is required that the germplasm materials preserved in long-term genebanks still have a high regeneration rate and maintain the original genetic traits when they are re-propagated after several years. [0003] Wasabi ( Eutrema wasabi (Sieboid) Maxim), also known as Behenia, is a perennial herb of the genus Behenia in the family Brassicaceae. It is native to East Asia and distributed in Southwest my country and Zhejiang provinces. Roots, stems and leaves all have extremely high economic value. The whole plant of wasabi has fragrant, pungent, sweet, sticky flavors and bactericidal properties. Its unique spicy taste makes it used as a unique condiment for food, and the leave...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N3/00A01H4/00
Inventor 陈晓玲张金梅辛霞卢新雄
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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