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Preparation method of tuber pinellia polyploid plant

A polyploid and Pinellia technology, applied in the field of plant cell engineering, can solve problems such as easy generation of chimeras, increased workload and complexity of mutant plant screening, and reverse mutation.

Inactive Publication Date: 2012-05-16
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Germinated seeds, seedlings, buds, callus and shoot apex tissues are often used as colchicine treatment materials for polyploid induction by such methods, and the treatment methods include dipping, smearing, dripping and injection, etc. Chromosome doubling induction is easily disturbed by the physiological conditions of the processed materials, and chimeras are prone to occur, and back mutations may occur, thereby increasing the workload and complexity of mutant plant screening

Method used

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  • Preparation method of tuber pinellia polyploid plant
  • Preparation method of tuber pinellia polyploid plant
  • Preparation method of tuber pinellia polyploid plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1. A preparation method of a peach leaf type Pinellia polyploid plant, followed by the following steps:

[0065] 1). Soak the young peach leaf-shaped pinellia leaves with 75% alcohol by volume for 30~45s, and then use HgCl with a mass concentration of 0.1% 2 Disinfect the solution for 1~5min, rinse with sterile water for 5~10 times; cut the sterilized leaves into 0.5cm 2 The small pieces of left and right were inoculated into callus induction medium and cultured for 14-16 days, the light time was 12h / d, the light intensity was 1500lx-2000lx, and the culture temperature was 22-28°C to obtain yellow-green hard callus;

[0066] The formula of callus induction medium is MS medium +6-BA 1.0mg·L -1 +NAA 0.02mg·L -1 +Sucrose 30g·L -1 , PH 5.6~6.0.

[0067] 2) Transfer the hard-textured callus to the callus subculture medium for light culture, the light time is 12h / d, the light intensity is 1500lx~2000lx, the culture temperature is 22~28℃; every 7~10d Subculture once, subcultu...

Embodiment 2

[0086] Example 2. Peach leaf type Pinellia was used as material for polyploid induction. The specific steps are as shown in Example 1. The difference is that the mass concentration of colchicine in step 5 is 0.003%, and the treatment time is 48 hours. The polyploid traits of is consistent with Example 1, and the polyploid induction rate of this treatment is 31.43%.

Embodiment 3

[0087] Example 3. Peach leaf type Pinellia was used as material for polyploid induction. The specific steps are as shown in Example 1. The difference is that the mass concentration of colchicine in step 5 is 0.005%, and the treatment time is 24 hours. The polyploid traits of is consistent with Example 1, and the polyploid induction rate of this treatment is 33.15%.

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Abstract

The invention discloses a preparation method of a tuber pinellia polyploid plant. In the method, tuber pinellia leaves are used for inducing callus with loose texture, then the callus is treated by suspension culture, tuber pinellia single cell is separated out to be processed by colchicines, then the tuber pinellia single cell is cultured in a tuber pinellia single cell culture medium after the colchicines is washed away, the formed cell mass is inoculated into the callus inducing culture medium for culturing, the formed callus is moved into a cluster buds culture medium for culturing and then moved into a rooting medium for rooting medium after the plant leaves are completed spread, and the root tips are taken for chromosome observation for confirming the ploidy of the plant, so as to obtain the tuber pinellia polyploid plant. The tuber pinellia polyploid plant obtained by the method has obvious polyploidy feature and the screening process is relatively simple.

Description

Technical field [0001] The invention belongs to the field of plant cell engineering, and specifically relates to a method for inducing polyploidy of Pinellia ternata, a medicinal plant, which uses a combination of plant tissue culture technology and cell suspension culture to induce polyploidy of Pinellia ternata. Background technique [0002] Pinellia (Pinellia spp.) is a plant of the genus Pinellia of the Araceae family, which mainly reproduces asexually. Dry tubers are a common traditional Chinese medicine. Modern pharmacological studies have shown that Pinellia has many effects such as drying dampness and reducing phlegm, reducing nibble and relieving cough, eliminating pimples, anti-early pregnancy, anti-tumor effect, lowering blood fat, protecting liver and treating coronary heart disease. At present, wild pinellia resources are gradually decreasing due to people's blind mining, and the genetic background and ploidy of artificially cultivated pinellia are complex and change...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01H4/00A01H1/08
Inventor 陈集双郭巧萍
Owner NANJING TECH UNIV
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