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Method for cultivating autotetraploid plants of stevia rebaudiana

A technology of autotetraploid and breeding method, which is applied in the cultivation of new plant varieties and the cultivation of sugar crop stevia autotetraploid plants, can solve the problem of low steviol glycoside content, achieve short internodes, Improve work efficiency, the effect of deep leaf color

Inactive Publication Date: 2012-08-01
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem of low content of steviol glycosides in the existing Stevia rebaudiana. According to the doubling of plant cell chromosomes, the doubling of its gene dose can enhance the metabolism of plants, and the content of effective chemical components for gene regulation on chromosomes is often higher than that of diploids. At the same time, polyploid plants have the characteristics of large organs, so a method for cultivating stevia autotetraploid plants is provided, which can cultivate stevia autotetraploid plants with higher content of active ingredients (steviol glycosides) in a short period of time to meet people's demand for steviol glycoside sweetness ingredients in the food and pharmaceutical industries

Method used

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  • Method for cultivating autotetraploid plants of stevia rebaudiana
  • Method for cultivating autotetraploid plants of stevia rebaudiana
  • Method for cultivating autotetraploid plants of stevia rebaudiana

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Experimental program
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Effect test

Embodiment 1

[0049] (1) Acquisition of adventitious buds: select the robust tender shoots of stevia plants, wash them with washing powder first, then rinse them with flowing water for 2 hours, soak them in 75% alcohol for 30 seconds, rinse them with sterile water for 2 to 3 times, and then put them in 0.10 % mercuric chloride solution for 5 minutes, rinsed with sterile water for 3 to 4 times, and inoculated the sterilized shoots into the following induction medium:

[0050] MS medium

[0051]

[0052] The cultivation room uses fluorescent lamps as the light source, the illuminance is 1500Lx, the illumination is 12h per day, and the temperature is (24±1)°C for 20 days to induce a large number of adventitious buds.

[0053] (2) Stevia autotetraploid induction: soak the adventitious buds in a 0.5g / L colchicine solution (adding 2% dimethyl sulfoxide) for 12-48 hours, soak 30 buds for each treatment Adventitious buds. After soaking, the adventitious buds were rinsed with sterile water for ...

Embodiment 2

[0063] (1) Acquisition of adventitious buds: select healthy young buds of Stevia rebaudiana plants, wash them with washing powder and then rinse them with flowing water for 1.5 hours. Soak in 75% alcohol for 30 seconds, rinse with sterile water for 2 to 3 times, then soak in 0.10% mercuric chloride solution for 6 minutes, and rinse with sterile water for 3 to 4 times. Inoculate the sterilized stem segments into induction medium: MS medium + 0.2mg / L6-BA + 0.1mg / L KT + 0.01mg / LNAA + 6500mg / L agar + 30000mg / L sucrose, pH 5.6. The cultivation room uses fluorescent lamps as the light source, the illuminance is 1500-2000Lx, the light is 12h per day, and the temperature is (24±1)°C for 15 days to induce a large number of adventitious buds.

[0064] (2) Stevia autotetraploid induction: soak the adventitious buds in a colchicine solution with a concentration of 1.0 g / L (adding 2% dimethyl sulfoxide) for 12 to 48 hours, soaking 30 buds for each treatment Adventitious buds. After soaki...

Embodiment 3

[0070] (1) Acquisition of adventitious buds: select robust young buds of stevia plants, wash them with washing powder and then rinse them with running water for 1-2 hours. Soak in 75% alcohol for 30 seconds, rinse with sterile water for 2 to 3 times, then soak in 0.10% mercury chloride solution for 5 minutes, and rinse with sterile water for 3 to 4 times. Inoculate the sterilized stem segment into the following induction medium: MS medium + 0.3mg / L6-BA + 0.2mg / L KT + 0.05mg / LNAA + 6500mg / L agar + 30000mg / L sucrose, pH 5.8 induction variable bud. The cultivation room uses fluorescent lamps as the light source, the illuminance is 1500-2000Lx, the light is 12h per day, and the temperature is (24±1)°C for 20 days to induce a large number of adventitious buds.

[0071] (2) Stevia autotetraploid induction: soak the adventitious buds in a 2.0 g / L colchicine solution (adding 2% dimethyl sulfoxide) for 12 to 48 hours, soaking 30 buds for each treatment Adventitious buds. After soaki...

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Abstract

The invention discloses a method for cultivating autotetraploid plants of stevia rebaudiana. The method disclosed by the invention comprises the following steps of: taking adventitious buds of the stevia rebaudiana as a mutagenic material in an excised condition, and carrying out induced culture of tetraploids by adopting a method for combining a Colchicine solution with a tissue culture technology. By means of the method disclosed by the invention, higher inductivity can be obtained; a lot of tetraploid tissue culture seedlings can be obtained in a short time; compared with diploids, the obtained autotetraploid tissue culture seedling internodes of the stevia rebaudiana are short, dark in leaf colour and large and thick in leaf blade; and the advantages of polyploidy on morphology are initially embodied.

Description

technical field [0001] The invention relates to a method for cultivating a new plant variety, in particular to a method for cultivating a sugar crop stevia autotetraploid plant, and belongs to the field of biotechnology breeding. Background technique [0002] Stevia rebaudiana Bertoni (Stevia rebaudiana Bertoni) is a perennial herb of the genus Asteraceae of the dicotyledonous plant class Asteraceae. It is native to the Amanbai Mountains in Paraguay, South America. Local residents have been drinking it as sweet tea for a hundred years. It is a new type of sugar crop. The leaves of stevia rebaudiana contain steviol glycoside, a sweet component, which is 250-300 times sweeter than sucrose, non-toxic, high sweetness, low calorie and no side effects. Stevia has high economic value and was successfully introduced in my country in 1976. In recent years, conventional breeding has done many experiments for the improvement of stevia varieties. With the development of biotechnology, ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 向增旭李红
Owner NANJING AGRICULTURAL UNIVERSITY
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