Gentic transforming method of perennial rye grass

A technology of ryegrass and purpose, applied in the field of genetic transformation of perennial ryegrass, can solve the problems of few reports on genetic transformation of ryegrass, large variation of cloned regenerated plants, difficulty in establishing protoplasts and protoplast regeneration systems, etc.

Inactive Publication Date: 2005-08-24
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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Problems solved by technology

However, these methods have disadvantages that are difficult to overcome: such as the protoplast fusion method, the transformation efficiency is low, the establishment of protoplasts and protoplast regeneration systems is very difficult, and the clonal regenerated plants vary greatly; the gene gun method has relatively high transformation efficiency. , but the integration site and copy number of exogenous genes are difficult to control, and the expression level and genetic stability of exogenous genes are poor (Hansen G, Wright MS, 1999; Wang Guanlin et al., 1998)
[0006] At present, there are very few reports on the genetic transformation of ryegrass mediated by Agrobacterium tumefaciens at home and abroad

Method used

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  • Gentic transforming method of perennial rye grass
  • Gentic transforming method of perennial rye grass
  • Gentic transforming method of perennial rye grass

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Experimental program
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Embodiment Construction

[0028] 1. Materials

[0029]Perennial ryegrass tested materials mainly include Accent, Derby, Supreme Derby, Gold Medalist, Top Gun, Duofu, Emerald (Green Gem), purchased from the market.

[0030] Tobacco varieties, mature seeds of Honghua Dajinyuan, can be purchased from the market. Used to increase conversion efficiency.

[0031] Agrobacterium tumefaciens strain EHA105, resistant to the antibiotic rifampicin (Rif), carries the binary vector plasmid pCAMBIA-2301, the expression cassette of which is shown in figure 1 . Such as figure 1 As shown, the plasmid carries the selection gene neomycin phosphotransferase gene (npt II) and the reporter gene β-glucuronidase gene (gusA), and gusA contains two catalase introns, so that it cannot be expressed in Agrobacterium. These two genes are both activated by the cauliflower mosaic virus 35S promoter, can replicate and express in plant cells, and can be directly introduced into Agrobacterium to transform plants. This plasmid is m...

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Abstract

A transgenic method for perennial ryegrass includes such steps as providing the induced calli of mature embryo, infecting by the Agrobacterium tumefaciens with antibiotic resistant gene, and antibiotic screening to obtain resistance plant.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular, the invention relates to the genetic transformation of perennial ryegrass (Lolium perenne L.) mediated by Agrobacterium tumefaciens Background technique [0002] Plants of the genus Lolium L. widely used as turfgrass and grass are mainly perennial ryegrass and annual ryegrass, and there are various cultivars. The perennial ryegrass (Lolium perenne L.) was originally distributed in the temperate regions of Asia, Europe, North Atlantic islands and North Africa (Jauhar pp, 1993), and now it is planted all over the world, and it belongs to the imported cultivation in my country. It has strong trampling and cold resistance, but weak tolerance to drought and heat. The cultivation of new varieties of fine ryegrass plays a very significant role in improving the yield and quality of forage grass, improving the resistance to adverse environments, improving natural grasslands, establi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/00
Inventor 马欣荣刘华玲杨宏
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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