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Culture medium for cultivating transgenic plant

A basic culture medium and culture medium technology, applied in the field of culture medium for cultivating transgenic plants, can solve the problems of cumbersome operation and unsuitability for domestic application

Inactive Publication Date: 2013-09-25
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Philippe Vain of the John Innes Center in the United Kingdom established an efficient transformation system with the sequencing strain Bd21. This system uses MS medium as the basic medium and hygromycin as the plant selection marker. The operation is cumbersome and not suitable for domestic applications.

Method used

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  • Culture medium for cultivating transgenic plant
  • Culture medium for cultivating transgenic plant
  • Culture medium for cultivating transgenic plant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, the preparation of culture medium

[0051] 1. Callus induction medium

[0052] Add 2,4-D (2,4-dichlorophenoxyacetic acid), maltose, L-asparagine, inositol, hydrolyzed casein, proline, thiamine hydrochloride, agar to MS minimal medium , to obtain the callus induction medium, wherein, the final concentration of 2,4-D in the callus induction medium is 2.5mg / L, the final concentration of maltose in the callus induction medium is 30mg / L, L-day The final concentration of paraparagine in the callus induction medium is 150 mg / L, the final concentration of inositol in the callus induction medium is 0.2 g / L, and the final concentration of hydrolyzed casein in the callus induction medium is 0.5 g / L, the final concentration of proline in the callus induction medium is 700mg / L, the final concentration of thiamine hydrochloride in the callus induction medium is 1.5mg / L, and the agar is in the callus induction medium The final concentration of the solution is 7g / L, and...

Embodiment 2

[0062] Embodiment 2, the acquisition of transgenic Brachypodium distachyon

[0063] The flow chart of obtaining transgenic Brachypodium distachyon is as follows figure 1 As shown, among them, A is Brachypodium distachyon Bd21, B is Brachypodium distachyon Bd21 immature embryo, C is callus induction culture, D is subculture, E is exogenous DNA infecting callus block, F is the fluorescent callus block, G is the callus regenerated seedling, H is the positive regenerated seedling.

[0064] 1. Obtaining explants

[0065] Brachypodium distachyon (L.) P.Beauv) Bd21 (Wang Honggui, Wang Baoli, Lin Chentao, Fu Yongfu, Morphological Observation of Brachypodium distachyon (L.) P.Beauv), Northwest Agricultural Journal, 2007, Issue 6, Ersachyon Morphological observations of Pinegrass Bd21, publicly available from the Institute of Botany, Chinese Academy of Sciences).

[0066] Planting of Bd21: Germination: Put Brachypodium seeds on absorbent paper at room temperature for 3-5 days (depend...

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Abstract

The invention discloses a culture medium for cultivating a transgenic plant. The culture medium group comprises a callus induction culture medium, a subculture culture medium, a co-culture culture medium, a screening culture medium and a regeneration culture medium. Experiment results show that: with a transformation system and a transformation method of the present invention, average every 100 brachypodium distachyon immature embryos can produce about 103 positive transformation seedlings, wherein a positive seedling transformation rate achieves 103.4%, and is far higher than positive seedling transformation rates of the traditional and grain model plant rice, and the highest rice positive seedling transformation rate is 47.6% (Chinese Bulletin of Botany, 2008, 25(3),322-331). With the efficient agrobacterium transformation system of the brachypodium distachyon, the important technology means is provided for temperate zone and grain plant gene function researches.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a culture medium for cultivating transgenic plants. Background technique [0002] The scientific name of Brachypodium distachyon (L.) P.Beauv, also known as Purple False Brome, originated in the Mediterranean and the Middle East, and belongs to the genus Brachypodieae of the subfamily Pooideae. In phylogenetic evolution, it belongs to the same evolutionary branch as Temperate cereals and Meadow and Forage grasses. Because the Brachypodium plant is short (15-20cm), the genome is only 272Mb, self-pollination, short growth period (8-12 weeks), and easy cultivation, it has rapidly developed into a new temperate and cereal grain and grass in recent years. Plant-like model plants. In February 2010, The International Brachypodium Initiative (The International Brachypodium Initiative) has completed the whole genome sequencing analysis of Brachypodium distachydium Bd21 (http: / / www.brachypod...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N5/10A01H4/00C12N15/84A01H5/00
Inventor 漆小泉赵素珍
Owner INST OF BOTANY CHINESE ACAD OF SCI
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