Ultralow temperature storage and regeneration culture method of in vitro stem tip of Jerusalem artichoke

A technology of ultra-low temperature preservation and regeneration culture, which is applied in the field of ultra-low temperature storage and regeneration culture of Jerusalem artichoke in vitro shoot tips, can solve problems such as different procedures and technologies, and achieve the effect of restoring good growth.

Active Publication Date: 2014-09-03
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A variety of cryopreservation methods have been developed. However, for different species or even for different varieties of the same species, the appropriate cryopreservation methods and procedures may be different.
So far, no universal cryopreservation method has been established in the world, which is also one of the main bottlenecks in the practical application of cryopreservation technology

Method used

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  • Ultralow temperature storage and regeneration culture method of in vitro stem tip of Jerusalem artichoke
  • Ultralow temperature storage and regeneration culture method of in vitro stem tip of Jerusalem artichoke
  • Ultralow temperature storage and regeneration culture method of in vitro stem tip of Jerusalem artichoke

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 Jerusalem artichoke test-tube plantlet isolated stem tip droplet vitrification cryopreservation technology and plant regeneration culture method

[0030] Experimental materials: from the National Germplasm Bank Test Tube Seedling Bank, the germplasm number is '2009331028'.

[0031] The specific steps of the experimental method are as follows:

[0032] 1. Material acquisition:

[0033] 1) Plant Jerusalem artichoke tubers in flowerpots, wait until they grow to about 40cm, cut off the robust branches as explants, cut them into about 3cm bud stem segments, wash the surface stains with tap water, and use 10% NaClO solution After the surface was disinfected for 8 minutes and washed with sterile water for 3 to 5 times, the explants were inoculated into MS+15g / L sucrose+8g / L agar medium for cultivation.

[0034] 2) After the test-tube plantlets grow stably, transfer to MS+0.1mg / L GA 3 +15g / L sucrose+8g / L agar medium for expansion and culture. The culture temper...

Embodiment 2

[0043] Example 2 Effect of sucrose concentration in the pre-culture solution on cryopreservation of isolated shoot tips of Jerusalem artichoke

[0044] Only the sucrose concentration in the pre-culture solution was changed, and the rest of the steps were the same as in Example 1 to study the effect of the sucrose concentration in the pre-culture solution on cryopreservation of Jerusalem artichoke isolated shoot tips. The result is as figure 2 shown.

[0045] from figure 2 It can be seen that when the sucrose concentration is 0.2mol / L, the survival rate and regeneration rate are both about 50%. With the increase of sucrose concentration, the survival rate and regeneration rate reached the highest when the concentration was 0.4mol / L, which were 93% and 83% respectively. Excessively high sucrose concentration will reduce the survival rate and regeneration rate. When the sucrose concentration is 0.5mol / L, the survival rate and regeneration rate are only about 10%. The shoot...

Embodiment 3

[0046] Example 3 Effect of pre-cultivation time on cryopreservation of isolated shoot tips of Jerusalem artichoke

[0047] Only the pre-cultivation time was changed, and the rest of the steps were the same as in Example 1, to study the effect of the pre-culture time on the cryopreservation of Jerusalem artichoke isolated shoot tips. The result is as image 3 shown.

[0048] from image 3 It can be seen that pre-culture has a significant effect on improving the survival rate and regeneration rate of Jerusalem artichoke isolated shoot tips after cryopreservation. With the prolongation of pre-culture time, the survival rate and regeneration rate showed a trend of increasing first and then decreasing. The survival rate and regeneration rate of the shoot tips were only 6% after cryopreservation without pre-culture; the survival rate and regeneration rate reached the maximum after 3 days of pre-culture, which were 93% and 83% respectively; after 4 days of pre-culture, the surviva...

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Abstract

The invention provides an ultralow temperature storage and regeneration culture method of an in vitro stem tip of Jerusalem artichoke. The method comprises the following steps: obtaining materials; peeling to obtain the stem tip; pre-culturing saccharose; loading a loading liquid; carrying out dehydration treatment by using PVS2 (Plant Vitrification Solution) vitrification protective liquid; carrying out ultralow temperature cryopreservation; unfreezing and washing; and recovering regenerative culture. By adopting the ultralow temperature storage technology by a droplet vitrification method, the vitro stem tip of a test-tube plantlet of Jerusalem artichoke stored at the ultralow temperature is defrosted and recovered in cultivation and is directly regenerated to form a seedling. The method is a long-term safe, stable, reliable and simple and effective method for storing germplasm resources of Jerusalem artichoke, wherein the survival rate can reach over 90%, the regeneration rate can reach over 80% and the regenerated plant is good in recovery growth.

Description

technical field [0001] The invention relates to a method for ultra-low temperature preservation of plant germplasm resources, in particular to a method for ultra-low temperature preservation and regeneration of Jerusalem artichoke isolated shoot tips. Background technique [0002] Jerusalem artichoke (Helianthus tuberous L.), also known as 'Jingjiang' and 'Devil's Ginger', is a perennial herbaceous plant of the Asteraceae Sunflower genus. It is named for its resemblance to a chrysanthemum on the ground and a taro below. Native to North America, it is widely distributed in the United States, Canada, Mexico and other countries. It was introduced into my country through Europe in the 17th century, and is currently distributed in the three northeastern provinces, Inner Mongolia, Beijing, Hebei, Henan, Hubei, Hunan, Gansu, Qinghai, Zhejiang, Shaanxi, Xinjiang, Anhui, Ningxia, Jiangsu, Shandong, Shanxi and other provinces and cities. Jerusalem artichoke has strong stress resistan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N3/00A01H4/00
Inventor 陈晓玲韩丽张金梅卢新雄辛霞尹广鹍何娟娟
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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