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A kind of genetic transformation method of german iris

A genetic transformation method and iris technology, which are applied in the directions of biochemical equipment and methods, horticultural methods, and botanical equipment and methods to achieve the effect of strong differentiation and regeneration ability and good infection effect.

Active Publication Date: 2022-02-15
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At home and abroad, many studies have been carried out on German Iris hybrid breeding, tissue culture and rapid propagation, and cut flower preservation. However, there are few reports on the genetic improvement of German Iris by using genetic engineering technology.

Method used

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  • A kind of genetic transformation method of german iris
  • A kind of genetic transformation method of german iris
  • A kind of genetic transformation method of german iris

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The invention relates to a method for constructing a German iris genetic transformation system, materials and reagents used: german iris stem tip, Agrobacterium (EHA105), plasmid (PBI121); absolute ethanol, mercury chloride, MS medium, 2,4- Dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA), 6-benzylaminopurine (6-BA), rifampicin (Rif), acetosyringone (AS), geneticin ( G418), Timentin (Tim), sucrose and agar powder. Main equipment: light incubator, ultra-clean workbench, centrifuge, PCR amplification instrument.

[0025] The steps are as follows:

[0026] 1. Induction and proliferation of German iris callus: dig up 2-3 year old German iris plants from the nursery and bring them back to the laboratory, cut off the leaves and rhizomes, keep about 1cm from the shoot tip, and use 10% washing solution Soak for 10 minutes, then rinse with running water for 1 hour, move to an ultra-clean workbench, disinfect with 75% ethanol for 45 seconds, rinse with sterile...

Embodiment 2

[0036] Take well-grown stem tips, young tender ovaries, young tender flower buds, and root tips respectively as explants, and the remaining processing steps are the same as in Example 1, and the pollution rate and callus induction rate of the explants are counted respectively at 45 days , the state and color of the callus, the results are shown in Table 1.

[0037] Table 1 Effects of different explants on callus induction of Iris germanica

[0038]

[0039]

[0040] According to the analysis of the above data, the present invention uses the shoot tip of Iris germanica as the explant, which has the characteristics of a callus induction rate as high as 82%, good embryonicity, and easy-to-obtain materials.

Embodiment 3

[0042] Using the processing method in Example 1, the difference is the acetosyringone and OD of the Agrobacterium infection solution 600 The value, statistical positive rate, the results are shown in Table 2.

[0043] Table 2 Different concentrations of acetosyringone and OD 600 The effect of the value of on conversion performance

[0044]

[0045] The above-mentioned data is analyzed and known, the acetosyringone of agrobacterium infection solution among the present invention is 200mg / L and OD 600 When the value of 0.4, the positive rate is the highest.

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Abstract

The method of the invention discloses an Agrobacterium-mediated genetic transformation method for Iris germanica, belonging to the technical field of plant genetic transformation. The method mainly takes the shoot tip callus of Iris germanica as a transformation recipient, and obtains transgenic strains through processes such as Agrobacterium infection and transformation, selection of resistant callus, redifferentiation and PCR detection of resistant plants. The method provided by the invention has the advantages of simple operation and high genetic transformation rate, and has great theoretical significance and application promotion value for the genetic transformation, gene function research and creation of new germplasm of German Iris and other Iris plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic transformation system construction, and in particular relates to a genetic transformation method of German iris. Background technique [0002] German Iris (Iris germanica L.) is a perennial herb of the genus Iris in the Iridaceae family. It is a famous ornamental flower. In addition to the variable-colored hairs on the ribs of the petals, this series of varieties also have large flowers, strange flower shapes, bright colors, Evergreen in four seasons and other characteristics have huge development space and prospects in landscaping, potted plants and ground cover applications. At home and abroad, many studies have been carried out on the hybrid breeding of German Iris, tissue culture and rapid propagation, and preservation of cut flowers. However, there are few reports on the genetic improvement of German Iris by using genetic engineering technology. It is of great significance to deeply st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/00A01H6/00
CPCA01H4/00C12N15/8205
Inventor 刘清泉张永侠原海燕黄苏珍唐君王玮琳杨永恒徐晓洋
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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