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Sweet-potato isolated culture adventitious-root germination method and use thereof

A sweet potato and culture substrate technology, applied in the field of botany, can solve the problems of limited number of transgenic plants, low transformation efficiency, and inability to be applied to production

Inactive Publication Date: 2012-01-11
南京晶薯生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example: the lack of an effective genetic transformation system, the transformation efficiency is low, the number of transgenic plants obtained is limited, and cannot be applied to production; there is a strong dependence on the genotype, so far, only chestnut fragrance, new big purple, high-line 14. Jewel, Jonathan and a few non-main varieties in production have obtained transgenic plants. These varieties are not main varieties, and they cannot be used in production
In addition, many researchers often obtain regenerated roots of sweet potatoes after transgenics but cannot obtain transgenic plants.

Method used

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  • Sweet-potato isolated culture adventitious-root germination method and use thereof
  • Sweet-potato isolated culture adventitious-root germination method and use thereof
  • Sweet-potato isolated culture adventitious-root germination method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1 sweet potato adventitious root sprouting method

[0074] 1. Selection of sweet potato strains

[0075] The present inventor selected 32 sweet potato lines for adventitious root sprouting experiment, and the selected lines are listed in Table 1, and these lines are all routinely used in the art.

[0076] Table 1

[0077]

[0078] 2. Preparation of culture medium

[0079] The medium used is sweet potato tissue culture basal medium, the composition is MS basal medium plus 0.3mg / LVB1, and the sucrose concentration is increased to 30g / L. Among them, the formula of MS basic medium is: macroelement: 440mg / LCaCl 2 2H 2 O, 170mg / L KH 2 PO 4 , 1900mg / L KNO 3 , 370mg / L MgSO 4 ·7H 2 O, 1650mg / LNH 4 NO 3 ;Trace elements: 0.025mg / L CoCl 2 ·6H 2 O, 0.025mg / L CuSO 4 ·5H 2 O, 0.25mg / LNa 2 MoO 4 2H 2 O, 6.2 mg / L H 3 BO 3 , 0.83mg / L KI, 16.9mg / L MnSO 4 ·H 2 O, 8.6mg / LZnSO 4 ·7H 2 O; Iron salt: 37.25mg / L Na 2 EDTA, 27.85mg / L FeSO 4 ·7H 2 O; organi...

Embodiment 2

[0103] Germination comparison under different culture conditions of embodiment 2

[0104] Sushu No. 2 (B11) and Taishu No. 6 (B22) with good bud growth were selected as test objects to verify the bud growth under different culture conditions.

[0105] Culture condition 1:

[0106] The cultivation method is the same as that in Example 1. After the root system is obtained, the root system is spread upside down on the fresh basic medium of sweet potato tissue culture.

[0107] Culture condition 2:

[0108] Other culture methods are the same as in Example 1, except: after obtaining the root system, the root system is tiled non-upside down (that is, according to the direction of the root system in the original culture bottle) on fresh sweet potato tissue culture basic medium.

[0109] 2 months after the root system was tiled to the fresh medium, it was observed whether buds appeared, and the number of buds produced on the root system was counted. The test results of Sushu No. 2 ...

Embodiment 3

[0115] Embodiment 3 prepares transgenic sweet potato plant

[0116] Using a conventional method, the expression vector pCAMBIA1301 (purchased from CAMBIA, with its own GUS gene) was introduced into competent Agrobacterium, and positive clones were screened to obtain the Agrobacterium into which the vector was introduced.

[0117] By the dipping method, the sweet potato explants were soaked with the Agrobacterium introduced with the vector, and some tissues were infected and the explants expressing the GUS gene (shown in blue) were as follows: Figure 5 shown. The cells infiltrated with Agrobacterium were screened by resistance selection, some of them were embryogenic cells (differentiable), and the embryogenic cells were placed on sweet potato tissue culture basic medium supplemented with 1 mg / L ABA, and cultured for about 1 month , transferred them to ABA-free sweet potato tissue culture minimal medium, and these embryogenic cells could gradually grow into roots.

[0118] W...

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Abstract

The invention discloses a method for producing sweet potato buds by isolated culture of adventitious roots by sweet potatoes. The sweet potato buds can be conveniently obtained by the method of the invention, and can produce a great deal of sweet potato plants by asexual propagation. The method of the invention has significance to the regeneration culture of sweet potatoes, and opens up a new wayfor sweet potato detoxication and genetic transformation to avoid complex tissue culture process. In addition, the method of the invention has wide applicability, and is applicable to most of sweet potato varieties.

Description

technical field [0001] The invention belongs to the field of botany; more specifically, the invention relates to a method for in vitro cultivation of adventitious roots of sweet potato and its application. Background technique [0002] Sweet potato belongs to Convolvulaceae, sweet potato genus, sweet potato species, is a vine herb. Sweet potato plants can be divided into roots, stems, leaves, flowers, fruits, seeds and other parts. Sweet potato is an important food crop and an important raw material in the food processing industry, with rich nutritional value. Therefore, the research on sweet potato variety optimization, cultivation and regeneration technology is of great significance. [0003] At present, the regeneration of isolated sweet potato plants is mainly achieved through the pathway of somatic embryogenesis. This process is highly dependent on genotype, and is very complicated and time-consuming. Therefore, it is necessary to explore other ways to rapidly and ef...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00C12N5/04C12N15/82
Inventor 张鹏李海霞杨俊
Owner 南京晶薯生物科技有限公司
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