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White birch transgenosis method

A transgenic, white birch technology, applied in the field of plant genetic engineering, can solve the problems of low birch transgenic efficiency, long cycle, and difficulty in degerming, and achieve the effects of shortening the transgenic cycle, strengthening differentiation ability, and shortening the transformation cycle

Active Publication Date: 2016-08-17
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a birch transgenic method in order to solve the problems of low birch transgenic efficiency, difficult degerming and long cycle

Method used

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  • White birch transgenosis method

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Experimental program
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specific Embodiment approach 1

[0025] Specific embodiment one: a kind of white birch transgenic method of this embodiment is realized according to the following steps:

[0026] 1. At 7 o'clock in the afternoon, select birch seeds with a germination rate of more than 70% and put them into a beaker. The mature white birch seeds at the bottom of the sink and the end of germination are used for transgenic recipients;

[0027] 2. Put the seeds selected in step 1 in a sterile petri dish, add 50ml of alcohol with a volume concentration of 75% to soak for 30s in an ultra-clean workbench, discard the alcohol, and then add 50ml of 30% hydrogen peroxide for disinfection for 15 minutes , discard the hydrogen peroxide, then wash the seeds 3 times with sterile water, and place them in a plate with sterile water for later use;

[0028] 3. At 21:00 on the day before step 2, take the engineered Agrobacterium pProBpIAA10::LUC stored at -80°C, inoculate it into 30ml of LB culture medium with corresponding antibiotics, shake ...

specific Embodiment approach 2

[0039] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the sign of the end of germination in Step 1 means that the radicle penetrates the tissue around the embryo. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0040] Specific embodiment 3: The difference between this embodiment and specific embodiment 1 is that in step 1, place it in a water tank and wash it with running water for 43 hours. Other steps and parameters are the same as those in Embodiment 1.

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Abstract

The invention discloses a white birch transgenosis method, belongs to the field of plant gene engineering and aims at solving the problem that white birch transgenosis is low in efficiency, difficult in sterilization and long in period. The method includes: 1, selecting white birch seeds, soaking, and picking mature seeds which are fully expanded by absorbing, submerged at the bottom of a bottle and germinated completely as transgenosis receptors; 2, sterilizing; 3, preparing engineering bacterium liquid used for infection of the seed receptors; 4, longitudinally cutting the seeds, and dip-dyeing; 5, placing the seeds in a liquid callus induction medium for co-culture; 6, obtaining a transgenosis strain to complete through callus culture, differential culture, subculture and rooting culture. By using the method, culture of a white birch explant is not needed, so that transgenosis period is shortened; a liquid co-culture mode is adopted, so that repeated sterilization steps are omitted, and white birch transgenosis efficiency is improved; conversion efficiency is high, period is short, and the problem that white birch transgenosis is low in efficiency, difficult in sterilization and long in period is solved. The white birch stain is high in transplanting survival rate, and the method is simple to operate and easy to master.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to a highly efficient and rapid transgenic method mediated by Agrobacterium and using birch germinated seeds as receptors. Background technique [0002] Birch (Betula platyphylla Suk.) is one of the rare broad-leaved tree species in Northeast China. However, due to the long breeding cycle of birch and the slow progress of genetic improvement, traditional birch breeding techniques cannot design and transform traits according to the breeding objectives to meet the growing social needs. Therefore, it is imperative to carry out genetic engineering breeding research on birch. [0003] At present, the insect-resistant, drought-resistant and salt-tolerant transgenic white birch has been obtained by using the Agrobacterium-mediated method, but the genetic transformation efficiency is low, and the transformation cycle of the Agrobacterium tumefaciens-mediated transformati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/00
CPCA01H4/00C12N15/8205
Inventor 徐文娣姜静刘桂丰王楚韩锐
Owner NORTHEAST FORESTRY UNIVERSITY
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