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A genetic transformation method of Iris germanica

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

Active Publication Date: 2019-01-11
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 genetic transformation method of Iris germanica
  • A genetic transformation method of Iris germanica
  • A genetic transformation method of Iris germanica

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The present invention is a method for constructing genetic transformation system of German iris. The materials and reagents used are: German iris stem tip, Agrobacterium (EHA105), plasmid (PBI121); absolute ethanol, mercury, 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 multiplication of callus of German iris: pick 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 top and bottom of the stem tip, and use 10% washing solution Soak for 10 minutes, then rinse with running water for 1 hour, move to the ultra-clean workbench, disinfect with 75% ethanol for 45 secon...

Embodiment 2

[0036] Take the well-growing stem tip, young tender ovary, young tender flower bud, and root tip as explants. The rest of the treatment steps are the same as in Example 1. The pollution rate and callus induction rate of the explants were counted at 45 days. The state and color of the callus, the results are shown in Table 1.

[0037] Table 1 The effect of different explants on callus induction of German iris

[0038]

[0039]

[0040] Analysis of the above data shows that the present invention uses the stem tip of German iris as the explant, which has the characteristics of a callus induction rate as high as 82%, good embryonicity, and easy availability of materials.

Embodiment 3

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

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

[0044]

[0045] Analysis of the above data shows that the acetosyringone of the Agrobacterium infection solution in the present invention is 200 mg / L and OD 600 When the value of is 0.4, the positive rate is the highest.

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Abstract

The invention discloses a genetic transformation method of Iris germanica mediated by Agrobacterium tumefaciens, belonging to the technical field of plant genetic transformation. The transgenic plantsare obtained by Agrobacterium tumefaciens infection, screening of resistant calli, redifferentiation and PCR detection of resistant plants, using stem tip calli of Iris germanica as transgenic recipients. The method provided by the invention has the advantages of simple operation and high genetic transformation rate, and has great theoretical significance and application and popularization valuefor genetic transformation of Iris germanica and other Iris plants, gene function research and creation of new germplasm.

Description

Technical field [0001] The invention belongs to the technical field of construction of plant genetic transformation systems, and specifically relates to a genetic transformation method of German iris. Background technique [0002] German iris (Iris germanica L.) is a perennial perennial herb of the Iris family Iris. It is a famous ornamental flower. This series of species has large flowers, peculiar flower patterns, and gorgeous colors, except for the changeable hairs on the ribs of the petals. The four-season evergreen and other characteristics have huge development space and prospects in landscaping, potted plants and ground cover applications. Many researches have been carried out at home and abroad in the cross-breeding of German iris, tissue culture and rapid propagation, and preservation of cut flowers. At present, there are few reports on the genetic improvement of German iris using genetic engineering technology. It is of great significance to in-depth study of flower-re...

Claims

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Application Information

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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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