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 low direct differentiation rate and long time, and achieve the effects of improving transformation efficiency, inhibiting overgrowth and high differentiation frequency.

Inactive Publication Date: 2012-10-31
SOUTH CHINA UNIV OF TECH
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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] In the following, the present invention will be further described in conjunction with the embodiments.

[0044] Examples of medium used:

[0045] ①Callus induction / subculture medium

[0046] NBB medium: N 6 Large elements+B 5 Trace element+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] ②Agrobacterium 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-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] ⑤Select the 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 .

[0054] ⑥Predifferentiation medium

[0055]...

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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 an efficient regeneration and transformation system of rice Zhonghua 11. Background technique [0002] Rice (Oryza sativa) is the most widely distributed and one of the most important food crops in the world, and its genetic transformation has always received extensive attention. Indica rice (ssp.indca) and japonica rice (ssp.japonica) are the two main subspecies of cultivated rice, with obvious differences in genetic transformation. In general, the embryogenic callus induction system of indica rice is difficult to establish, and the cell division ability is weak. The callus grows slowly after being transferred to the selective medium, and it is difficult to differentiate into seedlings. Japonica rice varieties are easy to transform successfully, and the transformation frequency is high. Zhonghua 11 is a japonica rice variety and is one o...

Claims

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Application Information

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Patent Type & Authority Patents(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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