Method for building high-efficiency regenerating and transforming system of Oryza sativaL. subsp. japonica 11

A high-efficiency regeneration and rice technology, applied in the field of plant genetic engineering, can solve the problems of long time consumption and low direct differentiation rate, and achieve the effects of inhibiting overgrowth, improving transformation efficiency, and ensuring transformation efficiency

Inactive Publication Date: 2011-10-19
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct differentiation rate of japonica rice varieties is less than 10%, and it takes a long time

Method used

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  • Method for building high-efficiency regenerating and transforming system of Oryza sativaL. subsp. japonica 11
  • Method for building high-efficiency regenerating and transforming system of Oryza sativaL. subsp. japonica 11
  • Method for building high-efficiency regenerating and transforming system of Oryza sativaL. subsp. japonica 11

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Embodiment Construction

[0043] The present invention will be further described below in conjunction with embodiment.

[0044] Example medium used:

[0045] ①Callus induction / subculture medium

[0046] NBB Medium: N 6 Mass elements + B 5 Trace elements + B 5 Organic ingredients + 2mg / L 2,4-D + 0.3g / l hydrolyzed casein + 0.5g / l proline + 30g / l sucrose + 3.5g / l vegetable gel, pH 5.8.

[0047] ② Agrobacterium activation medium

[0048] AB solid medium: AB medium + 5g / L glucose + 15g / L agar + 50mg / L rifampicin + 75mg / L kanamycin, adjust the pH to 7.2 with AB buffer.

[0049] ③AAI liquid medium: AAI medium+30g / l sucrose+70g / L glucose+2mg / L 2,4-D+200μMAS, pH 5.2.

[0050] ④Co-culture medium

[0051] NBB medium+0.3g / L hydrolyzed casein+0.5g / L proline+2mg / L 2,4-D+200μM AS+10g / L glucose+30g / L sucrose+3.5g / L plant gel, pH 5.5.

[0052] ⑤ Select culture medium

[0053] NBB+1g / L hydrolyzed casein+1g / L proline+2mg / L 2,4-D+sucrose+30mg / L hygromycin+500mg / L cephalosporin+3.5g / l vegetable gel, PH 5.8 .

...

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Abstract

The invention belongs to the field of genetic engineering of plants and discloses a method for building a high-efficiency regenerating and transforming system of Oryza sativaL. subsp. japonica 11 through cultivation of induction of embryogenic callus, subculture multiplication of embryogenic callus, transformation mediated by agrobacterium tumefaciens, cocultivation of embryogenic callus and agrobacterium tumefaciens, screening and cultivation of resistant callus and seedlings differentiation of resistant callus. The method has the advantages of simple technological processes, low production cost, and capabilities of obviously increasing the inductivity of the embryogenic callus, the subculture multiplication rate and the differentiation and regeneration rate of the embryogenic callus of the Oryza sativaL. subsp. japonica 11 and simultaneously shortening the period of obtaining regenerative rice plants; and on the basis, an improved agrobacterium tumefaciens-mediated method can be used for remarkably increasing the efficiency of obtaining the transgenosis rice plants and shortening the period of obtaining regenerative rice plants.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and specifically relates to a method for establishing a high-efficiency regeneration and transformation system of rice Zhonghua 11. Background technique [0002] Rice (Oryza sativa) is one of the most widely distributed and most important food crops in the world, and its genetic transformation has been widely valued. Indica rice (ssp.indca) and japonica rice (ssp.japonica) are the two main subspecies of rice cultivar, with obvious differences in genetic transformation. In general, the embryogenic callus induction system of indica rice is difficult to establish, the cell division ability is weak, and the callus grows slowly after being transferred to the selection medium, and it is difficult to differentiate into seedlings. Japonica rice varieties are easy to be successfully transformed, and the transformation frequency is high. Zhonghua 11 is a japonica rice variety, which is one of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N15/84A01H5/00
Inventor 王亚琴陈春峰陈兴瑶
Owner SOUTH CHINA UNIV OF TECH
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