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Method for culturing agrobacterium-mediated transgenic salix matsudana plants

An Agrobacterium-mediated, transgenic technology, applied in botany equipment and methods, horticultural methods, genetic engineering, etc., can solve the problems of inability to induce resistant buds, inability to obtain willow transgenic plants, etc., and achieve short induction time and high transformation rate High, short incubation time effect

Inactive Publication Date: 2013-04-24
NORTHEAST FORESTRY UNIVERSITY
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  • Description
  • Claims
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Benefits of technology

This patented technology can help create genetically modified plants that are more durable than traditional crops or trees by introducing resistance genes into their roots instead of just growing them out overnight at once. These induced germ cells have been found faster but with longer periods compared to previous methods such as cuttings or rootlets. They also require less space during production while still maintaining good quality grain due to its shorter cell division process. Overall this innovation makes plant growth quicker without compromising on yield potential.

Problems solved by technology

The technical problem addressed in this patents relates to improving the ability of tomatoes or other vegetables grown indoors during winters without causing harmful effects such as excessive greenhouse gases emissions that contribute to global warming issues. Current methods involve developing root systems with stronger biosynthetic capabilities but lack efficient ways to transform them back into sturdy ones like fertilizers due to their poor absorbance properties towards heavy metallics and organisms present within the environment.

Method used

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  • Method for culturing agrobacterium-mediated transgenic salix matsudana plants
  • Method for culturing agrobacterium-mediated transgenic salix matsudana plants
  • Method for culturing agrobacterium-mediated transgenic salix matsudana plants

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specific Embodiment approach 1

[0020] Specific embodiment one: the method for cultivating transgenic Salix sativa plant mediated by Agrobacterium in the present embodiment is carried out according to the following steps:

[0021] 1. Sterilize the mature willow seeds, then inoculate the sterilized seeds in the pre-cultivation medium and cultivate them for 4-8 days to obtain germinated seeds;

[0022] 2. Pick a single Agrobacterium tumefaciens colony carrying the pBI121 vector, insert it into LB liquid medium containing 50mg / L kanamycin, and culture it overnight at 28°C to obtain OD 600 0.2-1.2 Agrobacterium liquid;

[0023] 3. The germinated seeds obtained by pre-cultivation in step 1 are excised from the base to germinate the terminal bud position, and 2 cotyledons are reserved, and the germinated seeds with excised terminal buds are used as explants, and the explants are placed in step 2 for preparation OD 600 Soak in 0.2-1.2 Agrobacterium bacteria solution for 30-40 minutes;

[0024] 4. Then the explan...

specific Embodiment approach 2

[0030] Specific embodiment two: This embodiment is a further description of the pre-culture medium in step one of the method for cultivating transgenic Salix salix plants mediated by Agrobacterium described in specific embodiment one, and the pre-culture medium described in step one is MS medium containing 1.0 mg / L zeatin, 0.1 mg / L naphthaleneacetic acid, 30 g / L sucrose and 8 g / L agar.

specific Embodiment approach 3

[0031] Specific embodiment three: This embodiment is a further description of the method for disinfecting mature willow seeds in step one of the method for cultivating transgenic willow plants mediated by Agrobacterium described in specific embodiment one. The method for disinfecting mature willow seeds is specifically: soaking mature willow seeds in an ethanol solution with a volume concentration of 70% for 30 seconds in an ultra-clean workbench, rinsing with distilled water twice, and then transferring them to a 1% volume concentration Soak in sodium hypochlorite solution for 10 minutes, rinse with distilled water 5 times.

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Abstract

The invention discloses a method for culturing agrobacterium-mediated transgenic salix matsudana plants and relates to the method for culturing the transgenic salix matsudana plants. The method aims to solve the problems that the existing method cannot induce resistant buds from transgenic transgenices of salix matsudana so that the transgenic salix matsudana plants cannot be obtained. The method comprises the following steps of: firstly, sterilizing and preculturing salix matsudana seeds; secondly, selecting agrobacterium tumefaciens with pBI121 carriers, and culturing to obtain a bacteria solution; thirdly, taking sprouted top bud parts cut from basic parts as explants, and then soaking in the bacteria solution; fourthly, taking out the explants and soaking up through filter papers, and inoculating the explants into a co-culture culture medium; fifthly, switching to a selective medium till the resistant buds grow, switching to a light condition, and screening the resistant buds; sixthly, transferring the resistant buds into an elongation culture medium; seventhly, cutting off stem sections, and switching to a rooting culture medium for culture till roots grow; and finally, carrying out polymerase chain reaction (PCR) and glucuronidase (GUS) staining detection, transplanting to earth, and obtaining the transgenic salix matsudana plants. The method for culturing the agrobacterium-mediated transgenic salix matsudana plants is applied to culturing the transgenic salix matsudana plants.

Description

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Claims

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Application Information

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Owner NORTHEAST FORESTRY UNIVERSITY
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